{"gene":"PFKP","run_date":"2026-06-10T06:43:35","timeline":{"discoveries":[{"year":1983,"finding":"The PFKP locus was mapped to the short arm of human chromosome 10 (10p) using human X rodent somatic cell hybrids with active-enzyme immunoprecipitation; gene dosage effects were confirmed in fibroblasts with 10p duplication showing ~180% normal PFK activity.","method":"Somatic cell hybrid panel, active-enzyme immunoprecipitation with rodent anti-human P subunit-specific antiserum, gene dosage analysis","journal":"Human genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — nine of ten concordant hybrids, gene dosage experiment provides orthogonal confirmation, foundational mapping study","pmids":["6222962"],"is_preprint":false},{"year":2011,"finding":"KLF4 transcriptionally activates PFKP by directly binding the PFKP promoter in breast cancer cells, increasing glucose uptake and lactate production; knockdown of KLF4 reduced these glycolytic outputs specifically for the PFKP isoform without affecting other PFK isoforms.","method":"Promoter binding (ChIP/reporter assay), KLF4 knockdown and overexpression, glucose/lactate metabolic assays","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct promoter binding demonstrated, functional metabolic readouts, single lab","pmids":["21586797"],"is_preprint":false},{"year":2017,"finding":"Snail (SNAI1) transcriptionally represses PFKP in cancer cells undergoing EMT, diverting glucose flux from glycolysis toward the pentose phosphate pathway (PPP) and generating NADPH; knockdown of PFKP rescues the metabolic reprogramming and cell death induced by loss of Snail.","method":"Snail knockdown/overexpression, PFKP knockdown/rescue, metabolic flux assays (NADPH, PPP metabolites), in vivo metastasis assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal genetic epistasis (Snail loss rescued by PFKP knockdown), multiple orthogonal metabolic readouts, in vivo validation","pmids":["28176759"],"is_preprint":false},{"year":2018,"finding":"VDAC2 (a mitochondrial outer membrane protein) couples with PFKP on mitochondria to inhibit PFKP-mediated glycolysis; disruption of VDAC2 de-represses PFKP activity and drives non-stem glioma cells toward a stem cell phenotype, an effect blocked by PFK inhibitor clotrimazole.","method":"Co-expression analysis, VDAC2 overexpression/knockdown, PFK inhibitor (clotrimazole), stem cell marker assays, tumorigenicity assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — protein interaction inferred from co-localization and inhibitor rescue, single lab, multiple functional readouts","pmids":["30250190"],"is_preprint":false},{"year":2019,"finding":"Under hypoxia, oxidized ATM (DNA damage-independent activation) promotes upregulation of PFKP at the translational level via HIF1A, leading to intracellular citrate accumulation that enhances breast cancer cell migration and invasion through AKT/ERK/MMP2/9 signaling.","method":"ATM inhibition/knockdown, HIF1A regulation studies, PFKP protein/mRNA assays, citrate measurement, invasion/migration assays, xenograft experiments","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mechanistic dissection of ATM→HIF1A→PFKP axis, translational regulation established, single lab","pmids":["30850587"],"is_preprint":false},{"year":2021,"finding":"R-2-hydroxyglutarate (R-2HG) suppresses aerobic glycolysis in leukemia cells by inhibiting FTO-mediated m6A demethylation, which abrogates YTHDF2-dependent post-transcriptional upregulation of PFKP (and LDHB) mRNA; knockdown of FTO or PFKP phenocopies R-2HG-induced glycolytic inhibition, and PFKP overexpression reverses R-2HG effects.","method":"m6A sequencing, FTO/PFKP knockdown and overexpression, glycolysis assays, in vivo leukemogenesis models, primary AML cells","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (m6A-seq, genetic rescue, primary patient cells, in vivo), clear epistasis placing PFKP downstream of FTO/m6A/YTHDF2","pmids":["33434505"],"is_preprint":false},{"year":2021,"finding":"HRD1 (E3 ubiquitin ligase) interacts and co-localizes with PFKP in the cytoplasm, ubiquitinates PFKP targeting it for proteasomal degradation, and thereby reduces PFKP expression and enzymatic activity, suppressing aerobic glycolysis and breast cancer growth in a PFKP-dependent manner.","method":"Mass spectrometry, co-immunoprecipitation, immunofluorescence co-localization, ubiquitination assay, PFKP activity assay, in vivo xenograft","journal":"Cell communication and signaling : CCS","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, ubiquitination assay, MS identification of interaction, in vivo rescue experiments, multiple orthogonal methods","pmids":["33588886"],"is_preprint":false},{"year":2021,"finding":"YY1 transcription factor directly binds and transcriptionally activates the PFKP gene promoter in prostate cancer cells, forming an oncogenic YY1:BRD2/4-PFKP axis that sustains the Warburg effect; mutagenesis of YY1-bound cis-elements and gene loss-of-function/rescue studies confirmed the direct transcriptional regulation.","method":"Cistrome analysis (ChIP-seq), gene loss-of-function, YY1-bound cis-element mutagenesis, gene rescue studies, interactome profiling","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 2 / Strong — cistrome mapping, mutagenesis of binding elements, epistasis rescue experiments, multiple orthogonal approaches","pmids":["33849067"],"is_preprint":false},{"year":2021,"finding":"PFKP is a nucleocytoplasmic shuttling protein containing functional nuclear export and nuclear localization sequences (NLS). Cyclin D3/CDK6 promotes PFKP nuclear translocation by dimerization and by exposing the NLS, enabling interaction with importin 9. In the nucleus, PFKP stimulates CXCR4 expression via c-Myc activity to promote T-ALL cell invasion.","method":"NLS/NES mapping, Cyclin D3/CDK6 overexpression, importin 9 co-immunoprecipitation, PFKP nuclear fractionation, CXCR4 expression assays, in vivo leukemia homing assays, CXCR4 antagonist rescue","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 / Strong — nuclear localization signal mapping, protein-protein interaction (importin 9 Co-IP), c-Myc dependence, in vivo antagonist rescue, multiple orthogonal methods","pmids":["34255748"],"is_preprint":false},{"year":2021,"finding":"PFKP phosphorylates ATG4B at serine 34 (S34) in vitro, functioning as a protein kinase; amino acid deprivation strengthens the PFKP-ATG4B interaction, and this PFKP-mediated ATG4B phosphorylation enhances ATG4B activity and autophagic flux. PFKP S386 phosphorylation (under starvation) is required for ATG4B S34 phosphorylation.","method":"Tandem affinity purification-mass spectrometry, immunoprecipitation, in vitro kinase assay, CRISPR/Cas9 PFKP knockout, phosphosite mass spectrometry, autophagic flux assays","journal":"Cellular signalling","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro kinase assay directly demonstrating PFKP phosphorylates ATG4B-S34, confirmed by MS phosphosite mapping and CRISPR KO functional readout","pmids":["33607258"],"is_preprint":false},{"year":2021,"finding":"Mutant IDH1 upregulates PFKP expression through alteration of histone modification (as revealed by ChIP), and PFKP knockdown alleviates the mutant IDH1-induced increase in intrahepatic biliary organoid formation; high PFKP expression is more frequent in IDH-mutant cholangiocarcinoma.","method":"Gene expression analysis, ChIP for histone modification, Pfkp knockdown in intrahepatic biliary organoids, patient tissue immunohistochemistry","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP for epigenetic mechanism, organoid rescue experiment, single lab","pmids":["31827136"],"is_preprint":false},{"year":2022,"finding":"PFKP interacts with AMPK; upon glucose starvation this interaction is greatly enhanced and promotes mitochondrial recruitment of AMPK, which then phosphorylates ACC2 to enhance long-chain fatty acid oxidation, maintaining energy/redox homeostasis and promoting cancer cell survival.","method":"Proteomics screening of AMPK-interacting proteins, co-immunoprecipitation, PFKP knockdown/overexpression, ACC2 phosphorylation assays, fatty acid oxidation assays, metabolic measurements","journal":"Cell discovery","confidence":"High","confidence_rationale":"Tier 2 / Strong — proteomics-identified interaction validated by Co-IP, AMPK mitochondrial recruitment demonstrated, ACC2 phosphorylation as downstream readout, multiple orthogonal methods","pmids":["35641476"],"is_preprint":false},{"year":2022,"finding":"Fbxo7 promotes Cdk6-dependent phosphorylation of PFKP and Cdk6-independent ubiquitination of PFKP; loss of Fbxo7 reduces Cdk6 activity leading to increased glycolysis in CD4+ T cells, establishing PFKP as an essential Cdk6 substrate in T cells.","method":"Fbxo7 substrate screen, Fbxo7-deficient cells, metabolomics of activated CD4+ T cells, Cdk6 activity assays, ubiquitination assays, PFKP phosphorylation assays","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — substrate screen, Cdk6-dependent phosphorylation of PFKP validated, metabolomics confirmation, multiple orthogonal methods","pmids":["35670764"],"is_preprint":false},{"year":2022,"finding":"PFKP promotes accumulation of its product fructose-1,6-bisphosphate (FBP); FBP inhibits RhoA/ROCK1 pathway-mediated cytoskeletal remodeling in podocytes, and PFKP overexpression rescues podocytes from high-glucose-induced cytoskeletal remodeling via this FBP-dependent mechanism.","method":"AAV-mediated PFKP overexpression/knockdown in mouse kidneys, siRNA in PTECs, targeted metabolomics, FBP measurement, FBP exogenous addition, RhoA/ROCK1 pathway assays, aldolase B manipulation","journal":"Frontiers in endocrinology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — metabolomics and direct FBP addition experiments, RhoA/ROCK1 pathway readout, in vivo AAV model, single lab","pmids":["35095764"],"is_preprint":false},{"year":2023,"finding":"PFKP is lactylated at lysine 688 (K688), and this modification directly attenuates PFKP enzymatic activity, suggesting a negative feedback loop in glycolysis where lactate production leads to PFKP inhibition via lactylation.","method":"Mass spectrometry proteome-wide lactylation profiling, identification of PFKP K688 lactylation site, enzymatic activity assay of lactylated vs. non-lactylated PFKP","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — MS identification of modification site and in vitro enzymatic activity assay demonstrating inhibition, single lab","pmids":["38155775"],"is_preprint":false},{"year":2023,"finding":"CMBL (carboxymethylenebutenolidase-like) bridges TRIM25 E3 ubiquitin ligase to PFKP; ectopic CMBL enhances TRIM25 binding to PFKP, leading to PFKP ubiquitination and proteasomal degradation. p53 transcriptionally activates CMBL in response to genotoxic stress, repressing glycolysis via PFKP degradation.","method":"Co-immunoprecipitation (CMBL-TRIM25-PFKP ternary complex), ubiquitination assay, p53 transcriptional activation analysis, proteasome inhibition, CMBL/TRIM25 KO with PFKP rescue","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP of ternary complex, ubiquitination assay, genetic epistasis (p53→CMBL→TRIM25→PFKP), multiple orthogonal approaches","pmids":["37967006"],"is_preprint":false},{"year":2023,"finding":"TGF-β1 recruits the SMAD3-SP1 transcriptional complex to the PFKP promoter (confirmed by ChIP-qPCR) to enhance PFKP expression, upregulating glycolysis in renal proximal tubular epithelial cells and promoting kidney fibrosis; PFKP knockdown or overexpression in AAV-transduced mice correspondingly reduced or promoted fibrosis.","method":"ChIP-qPCR, AAV-mediated PFKP overexpression/knockdown in mouse kidneys, TGF-β1 stimulation of PTECs, glycolysis assays, fibrosis histology","journal":"Cell death & disease","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP-qPCR establishes direct SMAD3-SP1 binding to PFKP promoter, AAV in vivo functional validation, multiple orthogonal readouts","pmids":["38086793"],"is_preprint":false},{"year":2023,"finding":"SIRT2 deacetylates PFKP at mouse lysine 394 (human K395), impairing its glycolytic function and reducing ATG4B phosphorylation; reduced ATG4B activation leads to decreased LC3 activation, suppressed LC3-associated phagocytosis (LAP), and impaired pathogen clearance in ethanol-exposed macrophages.","method":"Bone marrow-derived macrophages from SIRT2-deficient mice, SIRT2 pharmacological inhibition, PFKP acetylation/deacetylation assays, ATG4B phosphorylation, LC3 activation assays, phagocytosis/LAP assays, bacterial clearance and sepsis survival","journal":"Frontiers in immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — specific modification site identified (K394/K395), genetic and pharmacological SIRT2 inhibition, multiple downstream functional readouts, in vivo survival data","pmids":["36865524"],"is_preprint":false},{"year":2023,"finding":"Glycolytic enzyme PFKP acts as a protein kinase for Lin41 in mouse embryonic stem cells: Pfkp phosphorylates Lin41 at serine residues, stabilizing Lin41 by impeding its autoubiquitination and proteasomal degradation; this allows Lin41 to destabilize ectodermal specification mRNAs and favor endodermal differentiation.","method":"Pfkp genetic manipulation (Stat3 repression model), in vitro/in vivo kinase assay for Lin41 phosphorylation, Lin41 ubiquitination assays, mESC differentiation lineage marker assays","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro kinase assay establishing PFKP as kinase for Lin41, ubiquitination/stability assays, lineage differentiation functional outcome, multiple orthogonal methods","pmids":["36660859"],"is_preprint":false},{"year":2023,"finding":"Triptolide (TP) inhibits PFKP expression in Sertoli cells through the GATA4/Sp1 transcriptional axis: GATA4 regulates Sp1, which drives PFKP expression; suppression of GATA4 or Sp1 reduces PFKP levels and impairs glycolysis.","method":"GATA4/Sp1 knockdown, Sp1 pharmacological inhibitor (plicamycin), PFKP mRNA/protein assays, glucose/lactate assays, in vivo mouse TP dosing","journal":"Toxicology and applied pharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic and pharmacological perturbation of transcriptional hierarchy, in vivo confirmation, single lab","pmids":["34087332"],"is_preprint":false},{"year":2023,"finding":"PFKP confers cisplatin resistance in NSCLC by upregulating the drug efflux transporter ABCC2 through activation of the NF-κB pathway (increased phospho-IκBα and nuclear p65); PFKP knockdown reduces NF-κB activation and ABCC2 expression, sensitizing cells to cisplatin.","method":"PFKP overexpression/knockdown in NSCLC cell lines, luciferase assay for ABCC2 promoter, NF-κB signaling assays (p-IκBα, nuclear p65), cisplatin sensitivity (flow cytometry), in vivo xenograft","journal":"Translational lung cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — luciferase assay linking PFKP to ABCC2 promoter via NF-κB, multiple functional assays, in vivo validation, single lab","pmids":["38090515"],"is_preprint":false},{"year":2024,"finding":"USP5 deubiquitinase directly binds PFKP and removes ubiquitin chains, stabilizing PFKP protein; USP5-mediated PFKP deubiquitination is essential for aerobic glycolysis and TNBC progression, with strong positive correlation between USP5 and PFKP levels in patient tissue.","method":"Co-immunoprecipitation, mass spectrometry protein identification, in vitro binding assay, ubiquitin assay, glycolysis assays, xenograft experiments, CPTAC database and IHC","journal":"Breast cancer research : BCR","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, in vitro binding assay, ubiquitination assay demonstrating deubiquitination, in vivo xenograft, MS identification, multiple orthogonal methods","pmids":["38217030"],"is_preprint":false},{"year":2024,"finding":"PFKP increases ERK-mediated stabilization of c-Myc protein, and c-Myc transcriptionally activates PFKP, forming a positive feedback loop that drives HNSCC progression; genetic and pharmacological co-targeting of PFKP and c-Myc shows synergistic anti-tumor effects in patient-derived organoids and xenografts.","method":"PFKP and c-Myc overexpression/knockdown, ERK signaling assays, c-Myc protein stability assays, c-Myc ChIP/transcriptional activation at PFKP, PDO and CDX/PDX models","journal":"Molecular cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis loop established by genetic perturbation and ChIP, multiple in vivo models, single lab","pmids":["38982480"],"is_preprint":false},{"year":2024,"finding":"RNF123 E3 ubiquitin ligase directly interacts with PFKP and induces its ubiquitination and degradation, thereby inhibiting glycolysis, cell cycle progression, and viability of breast cancer cells; overexpression of PFKP reverses the tumor-suppressive effects of RNF123.","method":"Co-immunoprecipitation, ubiquitination analysis, glycolysis assays, cell viability/cycle/colony formation assays, in vivo xenograft, lentiviral overexpression/silencing","journal":"Naunyn-Schmiedeberg's archives of pharmacology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP and ubiquitination assay, PFKP rescue experiment, single lab","pmids":["39725718"],"is_preprint":false},{"year":2024,"finding":"PFKP is lactylated at lysine 392 (K392); mutation of K392 diminishes PFKP lactylation and reduces glycolysis; PFKP K392 lactylation promotes glycolysis by regulating PTEN expression in hypoxia-treated ovarian cancer cells.","method":"Immunoprecipitation-western blot for PFKP lactylation, K392 site-directed mutagenesis, PFKP knockdown/overexpression, glycolysis assays, PTEN expression analysis, xenograft model","journal":"Biochemical genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — specific lactylation site mutagenesis, functional downstream PTEN regulation established, single lab","pmids":["39638933"],"is_preprint":false},{"year":2024,"finding":"PFKP binds AXL receptor tyrosine kinase and promotes its phosphorylation at Y779, activating AXL signaling and downstream MET phosphorylation in NSCLC cells, representing a non-metabolic oncogenic function of PFKP.","method":"PFKP knockdown (nanoparticle-mediated), co-immunoprecipitation of PFKP-AXL, AXL Y779 phosphorylation assays, MET phosphorylation assays, in vivo tumor growth","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP of PFKP-AXL interaction, phosphorylation readout at defined site, in vivo validation, single lab","pmids":["39664584"],"is_preprint":false},{"year":2024,"finding":"PFKP interacts with EIF2S2 (eukaryotic translation initiation factor 2 subunit beta); PFKP overexpression promotes new protein synthesis via EIF2S2, contributing to pathological cardiac hypertrophy; knockdown of EIF2S2 after PFKP overexpression reduces new protein synthesis and alleviates hypertrophy.","method":"Immunoprecipitation combined with LC-MS/MS to identify PFKP interactors, EIF2S2 knockdown rescue in NRCM and TAC mouse model, protein synthesis assays, cardiac phenotype characterization","journal":"Biochimica et biophysica acta. Molecular basis of disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MS-identified interaction, genetic rescue epistasis (EIF2S2 KD reverses PFKP overexpression phenotype), in vivo TAC model, single lab","pmids":["39419453"],"is_preprint":false},{"year":2025,"finding":"PKP1 (Plakophilin-1) stabilizes PFKP by binding TRIM21 and preventing TRIM21-mediated ubiquitination and proteasomal degradation of PFKP; PKP1 depletion selectively reduces PFKP levels, and functional rescue with PFKP restores the proliferative phenotype driven by PKP1.","method":"CRISPR knockout screening, PKP1 depletion, PFKP ubiquitination assays, TRIM21 binding assay, PFKP rescue experiments, metabolic assays (OCR/ECAR), multiple LUSC cell lines","journal":"Biomarker research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ubiquitination assay, TRIM21 binding, functional rescue, multiple cell lines, single lab","pmids":["40890861"],"is_preprint":false},{"year":2025,"finding":"HIF-1α transcriptionally controls PFKP mRNA in macrophages, as demonstrated by chromatin immunoprecipitation-qPCR (ChIP-qPCR); ethanol-induced oxidative stress impairs HIF-1α function, leading to reduced PFKP transcription, decreased glycolysis, and impaired phagocytosis; mitoquinol (MitoQ) restores HIF-1α function and PFKP expression.","method":"ChIP-qPCR for HIF-1α binding to PFKP promoter, macrophage ethanol exposure, PFKP mRNA/protein measurement, glycolysis assays, phagocytosis assays, MitoQ treatment, in vivo sepsis survival","journal":"Journal of immunology","confidence":"High","confidence_rationale":"Tier 2 / Strong — ChIP-qPCR directly demonstrates HIF-1α binding to PFKP promoter, pharmacological and genetic restoration of function, in vivo validation, orthogonal methods","pmids":["40356076"],"is_preprint":false},{"year":2025,"finding":"TRIS(2-chloroethyl)phosphate (TCEP) decreases PFKP glycolytic activity by enhancing O-linked N-acetylglucosamine (O-GlcNAc) transferase interaction with PFKP, thereby impairing glycolysis and ATP production in platelets, leading to suppression of platelet aggregation and activation.","method":"Proteomic analysis of platelets, O-GlcNAc transferase-PFKP interaction assay, PFKP activity assay, glycolysis/PPP pathway assays, ATP measurement, platelet activation assays, in vivo model","journal":"Environmental pollution","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — specific molecular interaction (OGT-PFKP) with PFKP activity readout, proteomics + functional assays, single lab","pmids":["39828204"],"is_preprint":false},{"year":2025,"finding":"Stat1 transcriptionally regulates Pfkp expression in macrophages; dsHMGB1 activates the Jak2/Stat1 pathway, which upregulates Pfkp to promote glycolysis and M1 macrophage polarization. ChIP-qPCR and dual-luciferase assays confirmed Stat1 binding to the Pfkp promoter.","method":"ChIP-qPCR, dual-luciferase assay, Jak2/Stat1 pathway inhibitor (fludarabine), dsHMGB1 treatment, glycolysis assays, macrophage polarization assays","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP-qPCR and luciferase confirm Stat1 binding to Pfkp promoter, pharmacological rescue, single lab","pmids":["41079919"],"is_preprint":false},{"year":2026,"finding":"ATM kinase phosphorylates PFKP at threonine 278 (T278) in response to ionizing radiation and high glucose, promoting PFKP transition from tetramers to dimers; nuclear dimeric PFKP recruits casein kinase 2 (CK2), which phosphorylates RAD51 at T13 to enhance RAD51-BRCA2 interaction and homologous recombination (HR) repair.","method":"In vitro phosphorylation assay (ATM kinase), phosphosite mutagenesis (T278), PFKP tetramer/dimer structural analysis, nuclear fractionation, CK2 recruitment Co-IP, RAD51 T13 phosphorylation assay, HR repair assay, irradiation and high glucose conditions","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro kinase assay, specific phosphosite mutagenesis, structural transition (tetramer→dimer) demonstrated, downstream CK2-RAD51-BRCA2 cascade validated by Co-IP and phosphorylation assay, multiple orthogonal methods","pmids":["42011782"],"is_preprint":false},{"year":2026,"finding":"PFKP directly binds AMOTL1 and inhibits its ubiquitin-mediated degradation; PFKP-driven aerobic glycolysis and EMT in HNSCC are AMOTL1-dependent. PFKP promotes YAP nuclear translocation via AMOTL1, suppressing Hippo pathway activity and amplifying glycolytic flux.","method":"Co-immunoprecipitation, ubiquitination analysis, AMOTL1 knockdown rescue, YAP nuclear translocation assays, Hippo pathway readouts, in vivo nude mice tumor models","journal":"Journal of translational internal medicine","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP, ubiquitination assay, AMOTL1 genetic rescue, in vivo confirmation, single lab","pmids":["41727965"],"is_preprint":false},{"year":2026,"finding":"USP14 deubiquitinase stabilizes PFKP through K48-linked deubiquitination; c-Myc transcriptionally upregulates USP14, while PFKP enhances ERK-dependent c-Myc protein stability, forming a c-Myc-USP14-PFKP feed-forward regulatory circuit that sustains PDAC chemoresistance.","method":"Ubiquitination assays (K48-linked), USP14-PFKP protein interaction studies, c-Myc transcriptional activation of USP14, ERK-c-Myc stability assays, patient-derived organoids, syngeneic and xenograft models, combined targeting experiments","journal":"Drug resistance updates","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — K48-specific ubiquitination assay, feed-forward circuit validated by multiple genetic/biochemical approaches, in vivo models, single lab","pmids":["41812332"],"is_preprint":false},{"year":2026,"finding":"PFKP K688 lactylation (K688la) is induced by hypoxia/ischemia and enhances PFKP enzymatic activity (distinct from K688 lactylation inhibiting activity in cancer cells [PMID 38155775]); a K688E mutation mimicking hyper-lactylation further amplifies glycolysis and confers cardiomyocyte cytoprotection.","method":"Lactylome proteomics (LAD-operated mice, hypoxic cardiomyocytes), PFKP K688E mutagenesis, PFKP enzymatic activity assays, ECAR/OCR metabolic assays, cardiomyocyte survival assays, 2-DG rescue","journal":"Frontiers in pharmacology","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — site-directed mutagenesis at K688 with enzymatic activity readout, in vivo and in vitro lactylome proteomics, single lab; note: contradicts PMID 38155775 on K688 lactylation effect direction","pmids":["41919225"],"is_preprint":false}],"current_model":"PFKP (phosphofructokinase, platelet isoform) is a rate-limiting glycolytic enzyme that catalyzes the phosphorylation of fructose-6-phosphate to fructose-1,6-bisphosphate, but it also functions as a non-canonical protein kinase (phosphorylating ATG4B-S34 to regulate autophagy and Lin41-serine to direct stem cell differentiation), undergoes nucleocytoplasmic shuttling (driven by Cyclin D3/CDK6-importin 9 interaction) to transcriptionally activate CXCR4 via c-Myc, and upon ATM-mediated T278 phosphorylation recruits CK2 to phosphorylate RAD51-T13 for homologous recombination repair; its activity and stability are regulated by multiple post-translational modifications including ubiquitination by HRD1, TRIM25/CMBL, RNF123, and USP14 (stabilized by USP5 and PKP1/TRIM21 competition), phosphorylation by CDK6 (via Fbxo7), deacetylation by SIRT2 at K394/K395 (reducing activity and LAP-mediated phagocytosis), and lactylation at K688 (with context-dependent activity effects); its transcription is activated by KLF4, YY1:BRD2/4, HIF-1α, Stat1, SMAD3-SP1, and c-Myc, and repressed by Snail, Stat3, and p53/CMBL, collectively placing PFKP at the intersection of glycolytic flux control, immune function, autophagy, DNA damage repair, and oncogenic signaling."},"narrative":{"mechanistic_narrative":"PFKP (platelet-type phosphofructokinase) is a rate-limiting glycolytic enzyme whose expression, stability, and moonlighting activities place it at the convergence of metabolic flux control, autophagy, DNA repair, and oncogenic signaling [PMID:28176759, PMID:33607258, PMID:42011782]. Its glycolytic output is controlled at multiple levels: a broad transcriptional network activates PFKP — KLF4 [PMID:21586797], YY1 with BRD2/4 [PMID:33849067], HIF-1α [PMID:40356076], Stat1 [PMID:41079919], SMAD3–SP1 [PMID:38086793], GATA4/Sp1 [PMID:34087332], and a c-Myc positive-feedback loop [PMID:38982480] — while Snail represses it to divert glucose into the pentose phosphate pathway [PMID:28176759], and p53 represses it indirectly by inducing CMBL [PMID:37967006]. PFKP protein stability is set by competing ubiquitin machineries, with the E3 ligases HRD1 [PMID:33588886], TRIM25 (bridged by CMBL) [PMID:37967006], RNF123 [PMID:39725718], and TRIM21 [PMID:40890861] driving proteasomal degradation, and the deubiquitinases USP5 [PMID:38217030] and USP14 [PMID:41812332] (the latter in a c-Myc–USP14–PFKP feed-forward circuit) stabilizing it. PFKP enzymatic activity is further tuned by post-translational modification, including SIRT2-mediated deacetylation at K394/K395 [PMID:36865524], lactylation at K688 and K392 [PMID:38155775, PMID:39638933, PMID:41919225], and O-GlcNAcylation [PMID:39828204]. Beyond catalysis, PFKP acts as a non-canonical protein kinase, phosphorylating ATG4B-S34 to promote autophagic flux [PMID:33607258] and Lin41 to stabilize it and direct endodermal differentiation in stem cells [PMID:36660859]. PFKP shuttles between cytoplasm and nucleus via mapped NLS/NES sequences, with Cyclin D3/CDK6 and importin 9 driving nuclear entry to transcriptionally activate CXCR4 through c-Myc [PMID:34255748]; upon ATM-mediated T278 phosphorylation it shifts from tetramer to dimer and recruits CK2 to phosphorylate RAD51-T13, supporting homologous recombination repair [PMID:42011782]. It also exerts protein-binding functions independent of catalysis, recruiting AMPK to mitochondria to drive fatty acid oxidation [PMID:35641476], activating AXL receptor signaling [PMID:39664584], and stabilizing AMOTL1 to promote YAP nuclear translocation [PMID:41727965].","teleology":[{"year":1983,"claim":"Establishing the chromosomal location of the platelet PFK subunit gene was the first step in defining PFKP as a distinct, dosage-sensitive genetic locus.","evidence":"Somatic cell hybrid panel with active-enzyme immunoprecipitation and gene-dosage analysis in fibroblasts","pmids":["6222962"],"confidence":"High","gaps":["Did not address tissue-specific regulation or non-glycolytic functions","No protein-level mechanism beyond enzyme dosage"]},{"year":2011,"claim":"Identifying KLF4 as a direct PFKP promoter activator showed PFKP transcription is selectively wired into oncogenic glycolytic programs distinct from other PFK isoforms.","evidence":"Promoter binding (ChIP/reporter), KLF4 knockdown/overexpression and glucose/lactate assays in breast cancer cells","pmids":["21586797"],"confidence":"Medium","gaps":["Single lab","Did not connect KLF4 to upstream signals controlling PFKP"]},{"year":2017,"claim":"Demonstrating Snail-mediated PFKP repression established PFKP as a metabolic switch directing glucose between glycolysis and the pentose phosphate pathway during EMT.","evidence":"Snail and PFKP knockdown/rescue with metabolic flux assays and in vivo metastasis models","pmids":["28176759"],"confidence":"High","gaps":["Mechanism of Snail recruitment to PFKP locus not detailed","PPP rerouting quantified but enzymatic regulation downstream of repression not addressed"]},{"year":2018,"claim":"Linking PFKP to VDAC2 at mitochondria revealed that spatial restriction of PFKP activity controls glioma stemness.","evidence":"Co-localization, VDAC2 manipulation, PFK inhibitor rescue and stem/tumorigenicity assays","pmids":["30250190"],"confidence":"Medium","gaps":["Interaction inferred from co-localization and inhibitor rescue rather than direct binding","Single lab"]},{"year":2019,"claim":"Defining an ATM→HIF1A→PFKP translational axis under hypoxia connected PFKP-derived citrate accumulation to invasive signaling.","evidence":"ATM inhibition/knockdown, HIF1A studies, citrate measurement and invasion/xenograft assays in breast cancer","pmids":["30850587"],"confidence":"Medium","gaps":["Translational regulation mechanism not molecularly resolved","Single lab"]},{"year":2019,"claim":"Mutant IDH1 was shown to upregulate PFKP via altered histone modification, tying oncometabolite-driven epigenetics to glycolytic gene expression.","evidence":"Expression analysis, ChIP for histone marks, organoid PFKP knockdown and patient IHC","pmids":["31827136"],"confidence":"Medium","gaps":["Specific histone mark and reader at PFKP locus not defined","Single lab"]},{"year":2021,"claim":"Multiple 2021 studies revealed PFKP is controlled post-transcriptionally (FTO/m6A/YTHDF2), by ubiquitin-mediated degradation (HRD1), and by direct transcriptional activation (YY1:BRD2/4), broadening the regulatory network governing PFKP abundance.","evidence":"m6A-seq with genetic rescue in leukemia; MS, reciprocal Co-IP and ubiquitination assays for HRD1; cistrome mapping and cis-element mutagenesis for YY1","pmids":["33434505","33588886","33849067"],"confidence":"High","gaps":["How these layers integrate to set net PFKP levels in a given cell not addressed","Each established in distinct cancer contexts"]},{"year":2021,"claim":"Discovery that PFKP phosphorylates ATG4B-S34 redefined PFKP as a non-canonical protein kinase coupling metabolic state to autophagy.","evidence":"TAP-MS, in vitro kinase assay, phosphosite MS and CRISPR knockout autophagic flux assays under amino acid deprivation","pmids":["33607258"],"confidence":"High","gaps":["Structural basis of PFKP kinase activity unresolved","Whether kinase and glycolytic activities are mutually exclusive not addressed"]},{"year":2021,"claim":"Mapping functional NLS/NES sequences and Cyclin D3/CDK6–importin 9-driven nuclear import established PFKP as a shuttling protein with a nuclear transcriptional function (CXCR4 via c-Myc).","evidence":"NLS/NES mapping, importin 9 Co-IP, nuclear fractionation, CXCR4 assays and in vivo homing/antagonist rescue in T-ALL","pmids":["34255748"],"confidence":"High","gaps":["Whether nuclear PFKP retains catalytic activity not resolved","Direct DNA contact vs c-Myc cofactor role not distinguished"]},{"year":2022,"claim":"Identifying PFKP–AMPK and CDK6–Fbxo7 axes showed PFKP both directs mitochondrial AMPK signaling under starvation and serves as an essential CDK6 substrate in T cells.","evidence":"Proteomics and Co-IP with ACC2 phosphorylation/FAO assays (AMPK); Fbxo7 substrate screen with metabolomics and ubiquitination/phosphorylation assays in CD4+ T cells (CDK6/Fbxo7)","pmids":["35641476","35670764"],"confidence":"High","gaps":["Functional residue(s) of CDK6 phosphorylation on PFKP not mapped","Crosstalk between PFKP scaffolding and catalysis not resolved"]},{"year":2022,"claim":"Demonstrating that PFKP product FBP inhibits RhoA/ROCK1 in podocytes linked PFKP catalytic output to cytoskeletal protection in a non-cancer setting.","evidence":"AAV PFKP manipulation in mouse kidney, targeted metabolomics, exogenous FBP addition and RhoA/ROCK1 readouts","pmids":["35095764"],"confidence":"Medium","gaps":["Direct FBP–RhoA/ROCK1 mechanism not biochemically defined","Single lab"]},{"year":2023,"claim":"A series of 2023 studies established the PTM and degradation logic of PFKP: lactylation (K688), deacetylation (K394/K395 by SIRT2), and CMBL-bridged TRIM25 ubiquitination under p53 control, each tuning enzymatic activity or stability.","evidence":"MS lactylation profiling with activity assays; SIRT2 KO/inhibition with ATG4B/LAP readouts; reciprocal Co-IP of CMBL–TRIM25–PFKP with ubiquitination and p53 epistasis","pmids":["38155775","36865524","37967006"],"confidence":"High","gaps":["Interplay among competing PTMs on the same protein not integrated","Stoichiometry and dynamics of modifications in vivo unclear"]},{"year":2023,"claim":"Identifying PFKP as a kinase for Lin41 extended its non-canonical kinase role to stem cell lineage control, stabilizing Lin41 to favor endodermal differentiation.","evidence":"Pfkp manipulation, in vitro/in vivo Lin41 kinase and ubiquitination assays, mESC lineage marker assays","pmids":["36660859"],"confidence":"High","gaps":["Lin41 target serine residues not fully enumerated","Relationship to ATG4B kinase activity not addressed"]},{"year":2023,"claim":"Adding SMAD3-SP1 (TGF-β1), GATA4/Sp1, and NF-κB–ABCC2 links expanded PFKP's roles to kidney fibrosis, Sertoli cell glycolysis, and NSCLC drug resistance.","evidence":"ChIP-qPCR with AAV in vivo fibrosis models; GATA4/Sp1 knockdown with glycolysis assays; PFKP–NF-κB–ABCC2 luciferase and cisplatin sensitivity assays","pmids":["38086793","34087332","38090515"],"confidence":"Medium","gaps":["Each axis from a single lab","Whether glycolytic vs moonlighting activity drives these phenotypes not always distinguished"]},{"year":2024,"claim":"2024 work deepened the PFKP stability and signaling map, adding USP5 and RNF123 to the ubiquitin axis, a c-Myc/ERK feedback loop, and lactylation at K392 regulating PTEN.","evidence":"Co-IP, in vitro binding and ubiquitination assays (USP5, RNF123); ERK/c-Myc stability and ChIP (HNSCC); K392 mutagenesis with PTEN/glycolysis readouts (ovarian cancer)","pmids":["38217030","39725718","38982480","39638933"],"confidence":"Medium","gaps":["Competing E3/DUB activities not reconciled into a unified stability model","Most findings single-lab and context-specific"]},{"year":2024,"claim":"Identifying PFKP binding partners AXL, EIF2S2, and AMOTL1 expanded its catalysis-independent scaffolding functions into receptor signaling, protein synthesis, and cardiac hypertrophy.","evidence":"Co-IP/MS with AXL Y779 and MET phosphorylation assays; LC-MS/MS with EIF2S2 rescue in TAC model; AMOTL1 binding/ubiquitination and YAP/Hippo assays","pmids":["39664584","39419453","41727965"],"confidence":"Medium","gaps":["Direct vs bridged interactions not always distinguished","Single-lab findings across distinct disease contexts"]},{"year":2025,"claim":"Showing HIF-1α, Stat1, and PKP1/TRIM21 each control PFKP refined macrophage and squamous-cancer regulation, including a PKP1-mediated block of TRIM21 ubiquitination.","evidence":"ChIP-qPCR and pharmacological rescue for HIF-1α and Stat1 in macrophages; CRISPR screening, TRIM21 binding and ubiquitination/rescue assays for PKP1 in LUSC","pmids":["40356076","41079919","40890861"],"confidence":"Medium","gaps":["O-GlcNAc and other PTM cross-regulation in immune cells not integrated","Most studies single-lab"]},{"year":2025,"claim":"Demonstrating that O-GlcNAc transferase interaction lowers PFKP activity in platelets connected PFKP-controlled glycolysis to platelet aggregation and a toxicant exposure phenotype.","evidence":"Platelet proteomics, OGT–PFKP interaction and activity assays with ATP/aggregation readouts","pmids":["39828204"],"confidence":"Medium","gaps":["O-GlcNAc site on PFKP not mapped","Single lab"]},{"year":2026,"claim":"Defining ATM–T278 phosphorylation that drives a tetramer-to-dimer transition and nuclear CK2 recruitment established PFKP as a direct participant in homologous recombination DNA repair.","evidence":"In vitro ATM kinase assay, T278 mutagenesis, oligomeric state analysis, CK2 Co-IP and RAD51-T13/HR repair assays under irradiation and high glucose","pmids":["42011782"],"confidence":"High","gaps":["Structural basis of dimer-specific CK2 recruitment not resolved","Whether dimeric PFKP retains glycolytic catalysis unclear"]},{"year":2026,"claim":"Adding a c-Myc–USP14–PFKP feed-forward circuit and a context-dependent K688 lactylation that enhances activity in cardiomyocytes refined PFKP stability control and reconciled tissue-specific PTM effects.","evidence":"K48-linked ubiquitination, USP14 interaction and ERK–c-Myc circuit in PDAC models; cardiac lactylome proteomics with K688E mutagenesis and activity/survival assays","pmids":["41812332","41919225"],"confidence":"Medium","gaps":["Opposite directionality of K688 lactylation effect between cancer and cardiomyocytes mechanistically unexplained","Single-lab findings"]},{"year":null,"claim":"How PFKP's catalytic, kinase, scaffolding, and nuclear functions are coordinately switched within a single cell, and what structural features distinguish its glycolytic from non-canonical activities, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model reconciling tetramer/dimer states with kinase and scaffold activities","Integration of competing E3/DUB and PTM inputs into net activity unknown","Context-dependent lactylation effects unexplained at the mechanistic level"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[9,18,31]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[9,18]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[8]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[6]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[8,31]},{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[3,11]}],"pathway":[{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[2,6,21]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[9,17]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[31]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[8]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[6,15,21,33]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[17,28,30]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[25,32]}],"complexes":["CMBL–TRIM25–PFKP ubiquitination complex"],"partners":["ATG4B","AMPK","VDAC2","AXL","AMOTL1","EIF2S2","TRIM25","USP5"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q01813","full_name":"ATP-dependent 6-phosphofructokinase, platelet type","aliases":["6-phosphofructokinase type C","Phosphofructo-1-kinase isozyme C","PFK-C","Phosphohexokinase"],"length_aa":784,"mass_kda":85.6,"function":"Catalyzes the phosphorylation of D-fructose 6-phosphate to fructose 1,6-bisphosphate by ATP, the first committing step of glycolysis","subcellular_location":"Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q01813/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/PFKP","classification":"Not Classified","n_dependent_lines":8,"n_total_lines":1208,"dependency_fraction":0.006622516556291391},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"HSP90B1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/PFKP","total_profiled":1310},"omim":[{"mim_id":"610681","title":"PHOSPHOFRUCTOKINASE, MUSCLE TYPE; PFKM","url":"https://www.omim.org/entry/610681"},{"mim_id":"603368","title":"CYCLIN-DEPENDENT KINASE 6; CDK6","url":"https://www.omim.org/entry/603368"},{"mim_id":"171860","title":"PHOSPHOFRUCTOKINASE, LIVER TYPE; PFKL","url":"https://www.omim.org/entry/171860"},{"mim_id":"171840","title":"PHOSPHOFRUCTOKINASE, PLATELET TYPE; PFKP","url":"https://www.omim.org/entry/171840"},{"mim_id":"123834","title":"CYCLIN D3; CCND3","url":"https://www.omim.org/entry/123834"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cytosol","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"retina","ntpm":250.0}],"url":"https://www.proteinatlas.org/search/PFKP"},"hgnc":{"alias_symbol":["PFK-C","PFKF"],"prev_symbol":[]},"alphafold":{"accession":"Q01813","domains":[{"cath_id":"3.40.50.450","chopping":"18-185_315-369","consensus_level":"high","plddt":93.4372,"start":18,"end":369},{"cath_id":"3.40.50.460","chopping":"188-311","consensus_level":"high","plddt":96.124,"start":188,"end":311},{"cath_id":"3.40.50.450","chopping":"406-539_686-736","consensus_level":"high","plddt":94.8501,"start":406,"end":736},{"cath_id":"3.40.50.460","chopping":"553-675","consensus_level":"medium","plddt":95.8321,"start":553,"end":675}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q01813","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q01813-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q01813-F1-predicted_aligned_error_v6.png","plddt_mean":92.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=PFKP","jax_strain_url":"https://www.jax.org/strain/search?query=PFKP"},"sequence":{"accession":"Q01813","fasta_url":"https://rest.uniprot.org/uniprotkb/Q01813.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q01813/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q01813"}},"corpus_meta":[{"pmid":"33434505","id":"PMC_33434505","title":"R-2-hydroxyglutarate 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metabolism.","date":"2020","source":"Cellular oncology (Dordrecht, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/32219704","citation_count":83,"is_preprint":false},{"pmid":"30850587","id":"PMC_30850587","title":"Intracellular citrate accumulation by oxidized ATM-mediated metabolism reprogramming via PFKP and CS enhances hypoxic breast cancer cell invasion and metastasis.","date":"2019","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/30850587","citation_count":76,"is_preprint":false},{"pmid":"35641476","id":"PMC_35641476","title":"PFKP alleviates glucose starvation-induced metabolic stress in lung cancer cells via AMPK-ACC2 dependent fatty acid oxidation.","date":"2022","source":"Cell discovery","url":"https://pubmed.ncbi.nlm.nih.gov/35641476","citation_count":73,"is_preprint":false},{"pmid":"29894707","id":"PMC_29894707","title":"Silencing PFKP inhibits starvation-induced autophagy, glycolysis, and epithelial mesenchymal transition in oral squamous cell 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biology","url":"https://pubmed.ncbi.nlm.nih.gov/35670764","citation_count":11,"is_preprint":false},{"pmid":"39638933","id":"PMC_39638933","title":"PFKP Lactylation Promotes the Ovarian Cancer Progression Through Targeting PTEN.","date":"2024","source":"Biochemical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/39638933","citation_count":11,"is_preprint":false},{"pmid":"33607258","id":"PMC_33607258","title":"PFKP facilitates ATG4B phosphorylation during amino acid deprivation-induced autophagy.","date":"2021","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/33607258","citation_count":10,"is_preprint":false},{"pmid":"38305210","id":"PMC_38305210","title":"Effect of lncRNA XIST on acute myeloid leukemia cells via miR-142-5p-PFKP axis.","date":"2024","source":"Hematology (Amsterdam, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/38305210","citation_count":10,"is_preprint":false},{"pmid":"36660859","id":"PMC_36660859","title":"Glycolytic Pfkp acts as a Lin41 protein kinase to promote endodermal differentiation of embryonic stem cells.","date":"2023","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/36660859","citation_count":8,"is_preprint":false},{"pmid":"39664584","id":"PMC_39664584","title":"PFKP silencing suppresses tumor growth via the AXL-MET axis.","date":"2024","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/39664584","citation_count":4,"is_preprint":false},{"pmid":"39419453","id":"PMC_39419453","title":"PFKP inhibition protects against pathological cardiac hypertrophy by regulating protein synthesis.","date":"2024","source":"Biochimica et biophysica acta. Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/39419453","citation_count":4,"is_preprint":false},{"pmid":"38044588","id":"PMC_38044588","title":"PFKP is upregulated in 5-fluorouracil-resistant patients and suppresses the antitumor activity of 5-fluorouracil in colorectal cancer in vitro and in vivo.","date":"2023","source":"Journal of chemotherapy (Florence, Italy)","url":"https://pubmed.ncbi.nlm.nih.gov/38044588","citation_count":4,"is_preprint":false},{"pmid":"39725718","id":"PMC_39725718","title":"RNF123 inhibits cell viability, cell cycle and colony formation of breast cancer by inhibiting glycolysis via ubiquitination of PFKP.","date":"2024","source":"Naunyn-Schmiedeberg's archives of pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/39725718","citation_count":4,"is_preprint":false},{"pmid":"40890861","id":"PMC_40890861","title":"PKP1 promotes lung cancer by modulating energy metabolism through stabilization of PFKP.","date":"2025","source":"Biomarker research","url":"https://pubmed.ncbi.nlm.nih.gov/40890861","citation_count":3,"is_preprint":false},{"pmid":"36110408","id":"PMC_36110408","title":"PFKP and GPC6 Variants Were Correlated with Alcohol-Induced Femoral Head Necrosis Risk in the Chinese Han Population.","date":"2022","source":"Pharmacogenomics and personalized medicine","url":"https://pubmed.ncbi.nlm.nih.gov/36110408","citation_count":3,"is_preprint":false},{"pmid":"40356076","id":"PMC_40356076","title":"Mitoquinol improves phagocytosis and glycolysis in ethanol-exposed macrophages via HIF-1α-PFKP axis.","date":"2025","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/40356076","citation_count":2,"is_preprint":false},{"pmid":"38427179","id":"PMC_38427179","title":"Silencing circLDLRAD3 Inhibits Lung Cancer Progression by Regulating the miR-497-5p/PFKP Axis.","date":"2024","source":"Molecular biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/38427179","citation_count":2,"is_preprint":false},{"pmid":"39828204","id":"PMC_39828204","title":"Inhibition of platelet activation process upon tris (2-chloroethyl) phosphate exposure: Role of PFKP-mediated glycolysis and the pentose phosphate pathway.","date":"2025","source":"Environmental pollution (Barking, Essex : 1987)","url":"https://pubmed.ncbi.nlm.nih.gov/39828204","citation_count":2,"is_preprint":false},{"pmid":"41079919","id":"PMC_41079919","title":"dsHMGB1, released from IL-17A-induced pyroptotic prostate epithelial cells, drives M1 polarization by promoting Pfkp-mediated glycolysis via Jak2/Stat1 transcription in experimental autoimmune prostatitis.","date":"2025","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/41079919","citation_count":1,"is_preprint":false},{"pmid":"40385478","id":"PMC_40385478","title":"Gen-miR-5 derived from Gentianella acuta inhibits PFKP to prevent fibroblast activation and alleviate myocardial fibrosis.","date":"2025","source":"Frontiers in pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/40385478","citation_count":1,"is_preprint":false},{"pmid":"40762768","id":"PMC_40762768","title":"P4HA2 promotes the progression of thyroid cancer by regulating PFKP-mediated glycolysis.","date":"2025","source":"Journal of physiology and biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/40762768","citation_count":1,"is_preprint":false},{"pmid":"41812332","id":"PMC_41812332","title":"The c-Myc-USP14-PFKP axis sustains chemoresistance in pancreatic ductal adenocarcinoma.","date":"2026","source":"Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy","url":"https://pubmed.ncbi.nlm.nih.gov/41812332","citation_count":0,"is_preprint":false},{"pmid":"40821334","id":"PMC_40821334","title":"PFKP Mediates Breast Cancer Metastasis Through Altered Glycolysis.","date":"2025","source":"Cureus","url":"https://pubmed.ncbi.nlm.nih.gov/40821334","citation_count":0,"is_preprint":false},{"pmid":"41448552","id":"PMC_41448552","title":"Integrative analysis of single-cell and bulk transcriptomes reveals metabolic heterogeneity and identifies PFKP as a therapeutic target in cervical cancer.","date":"2025","source":"Biochimica et biophysica acta. Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/41448552","citation_count":0,"is_preprint":false},{"pmid":"40651151","id":"PMC_40651151","title":"TRIM25 potentiates innate immune response to Senecavirus A by enhancing K63-linked ubiquitination of RIG-I and K48-linked ubiquitin-facilitated degradation of PFKP.","date":"2025","source":"Veterinary microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/40651151","citation_count":0,"is_preprint":false},{"pmid":"41727965","id":"PMC_41727965","title":"PFKP binding AMOTL1 promotes tumor aerobic glycolysis and epithelial-mesenchymal transition by modulating Hippo pathway in head and neck cancer.","date":"2026","source":"Journal of translational internal medicine","url":"https://pubmed.ncbi.nlm.nih.gov/41727965","citation_count":0,"is_preprint":false},{"pmid":"42011782","id":"PMC_42011782","title":"ATM promotes PFKP nuclear translocation to balance glycolysis and homologous recombination repair.","date":"2026","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/42011782","citation_count":0,"is_preprint":false},{"pmid":"42044203","id":"PMC_42044203","title":"PFKP Drives Immune Checkpoint Co-Expression and Metabolic Pathway Activation in Liver Cancer: TCGA-Based and Experimental Validation.","date":"2026","source":"Clinical and translational gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/42044203","citation_count":0,"is_preprint":false},{"pmid":"41919225","id":"PMC_41919225","title":"Lactylation of PFKP-K688 enhances glycolytic flux and confers cardioprotection in myocardial ischemia.","date":"2026","source":"Frontiers in pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/41919225","citation_count":0,"is_preprint":false},{"pmid":"42065039","id":"PMC_42065039","title":"Isoliquiritigenin Impedes Breast Cancer Progression through PITX1-PFKP-Mediated Glycolysis Reprogramming.","date":"2026","source":"Oncology research","url":"https://pubmed.ncbi.nlm.nih.gov/42065039","citation_count":0,"is_preprint":false},{"pmid":"42024199","id":"PMC_42024199","title":"PFKP is required for chemoresistant phenotype of breast cancer through modulating the formation of CD133+ cancer stem like cells.","date":"2026","source":"Molecular biomedicine","url":"https://pubmed.ncbi.nlm.nih.gov/42024199","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.03.19.25324051","title":"Multi-level Regulatory Roles of Lactate Metabolism Gene Network in Oral Cancer: Machine Learning Insights","date":"2025-03-20","source":"bioRxiv","url":"https://doi.org/10.1101/2025.03.19.25324051","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.05.22.655446","title":"Pneumococcal H  <sub>2</sub>  O  <sub>2</sub>  Reshapes Mitochondrial Function and Reprograms Host Cell Metabolism","date":"2025-05-22","source":"bioRxiv","url":"https://doi.org/10.1101/2025.05.22.655446","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.05.30.656396","title":"Molecular subtyping based on hippocampal cryptic exon burden reveals proteome-wide changes associated with TDP-43 pathology across the spectrum of LATE and Alzheimer’s Disease","date":"2025-06-03","source":"bioRxiv","url":"https://doi.org/10.1101/2025.05.30.656396","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.05.09.651611","title":"The Establishment of Prostate-specific, SKP2 Humanized Mice by CRISPR Knock-in Method Reveals Neoplastic Initiation and Microenvironmental Reprogramming","date":"2025-05-10","source":"bioRxiv","url":"https://doi.org/10.1101/2025.05.09.651611","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":35084,"output_tokens":9558,"usd":0.124311,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":19643,"output_tokens":6709,"usd":0.13297,"stage2_stop_reason":"end_turn"},"total_usd":0.257281,"stage1_batch_id":"msgbatch_01CNoDbkChHEkgrJnL7BwtGJ","stage2_batch_id":"msgbatch_016Y4x9E9Wg5VQDANj9quSQD","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1983,\n      \"finding\": \"The PFKP locus was mapped to the short arm of human chromosome 10 (10p) using human X rodent somatic cell hybrids with active-enzyme immunoprecipitation; gene dosage effects were confirmed in fibroblasts with 10p duplication showing ~180% normal PFK activity.\",\n      \"method\": \"Somatic cell hybrid panel, active-enzyme immunoprecipitation with rodent anti-human P subunit-specific antiserum, gene dosage analysis\",\n      \"journal\": \"Human genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — nine of ten concordant hybrids, gene dosage experiment provides orthogonal confirmation, foundational mapping study\",\n      \"pmids\": [\"6222962\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"KLF4 transcriptionally activates PFKP by directly binding the PFKP promoter in breast cancer cells, increasing glucose uptake and lactate production; knockdown of KLF4 reduced these glycolytic outputs specifically for the PFKP isoform without affecting other PFK isoforms.\",\n      \"method\": \"Promoter binding (ChIP/reporter assay), KLF4 knockdown and overexpression, glucose/lactate metabolic assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct promoter binding demonstrated, functional metabolic readouts, single lab\",\n      \"pmids\": [\"21586797\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Snail (SNAI1) transcriptionally represses PFKP in cancer cells undergoing EMT, diverting glucose flux from glycolysis toward the pentose phosphate pathway (PPP) and generating NADPH; knockdown of PFKP rescues the metabolic reprogramming and cell death induced by loss of Snail.\",\n      \"method\": \"Snail knockdown/overexpression, PFKP knockdown/rescue, metabolic flux assays (NADPH, PPP metabolites), in vivo metastasis assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal genetic epistasis (Snail loss rescued by PFKP knockdown), multiple orthogonal metabolic readouts, in vivo validation\",\n      \"pmids\": [\"28176759\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"VDAC2 (a mitochondrial outer membrane protein) couples with PFKP on mitochondria to inhibit PFKP-mediated glycolysis; disruption of VDAC2 de-represses PFKP activity and drives non-stem glioma cells toward a stem cell phenotype, an effect blocked by PFK inhibitor clotrimazole.\",\n      \"method\": \"Co-expression analysis, VDAC2 overexpression/knockdown, PFK inhibitor (clotrimazole), stem cell marker assays, tumorigenicity assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — protein interaction inferred from co-localization and inhibitor rescue, single lab, multiple functional readouts\",\n      \"pmids\": [\"30250190\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Under hypoxia, oxidized ATM (DNA damage-independent activation) promotes upregulation of PFKP at the translational level via HIF1A, leading to intracellular citrate accumulation that enhances breast cancer cell migration and invasion through AKT/ERK/MMP2/9 signaling.\",\n      \"method\": \"ATM inhibition/knockdown, HIF1A regulation studies, PFKP protein/mRNA assays, citrate measurement, invasion/migration assays, xenograft experiments\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mechanistic dissection of ATM→HIF1A→PFKP axis, translational regulation established, single lab\",\n      \"pmids\": [\"30850587\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"R-2-hydroxyglutarate (R-2HG) suppresses aerobic glycolysis in leukemia cells by inhibiting FTO-mediated m6A demethylation, which abrogates YTHDF2-dependent post-transcriptional upregulation of PFKP (and LDHB) mRNA; knockdown of FTO or PFKP phenocopies R-2HG-induced glycolytic inhibition, and PFKP overexpression reverses R-2HG effects.\",\n      \"method\": \"m6A sequencing, FTO/PFKP knockdown and overexpression, glycolysis assays, in vivo leukemogenesis models, primary AML cells\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (m6A-seq, genetic rescue, primary patient cells, in vivo), clear epistasis placing PFKP downstream of FTO/m6A/YTHDF2\",\n      \"pmids\": [\"33434505\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"HRD1 (E3 ubiquitin ligase) interacts and co-localizes with PFKP in the cytoplasm, ubiquitinates PFKP targeting it for proteasomal degradation, and thereby reduces PFKP expression and enzymatic activity, suppressing aerobic glycolysis and breast cancer growth in a PFKP-dependent manner.\",\n      \"method\": \"Mass spectrometry, co-immunoprecipitation, immunofluorescence co-localization, ubiquitination assay, PFKP activity assay, in vivo xenograft\",\n      \"journal\": \"Cell communication and signaling : CCS\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, ubiquitination assay, MS identification of interaction, in vivo rescue experiments, multiple orthogonal methods\",\n      \"pmids\": [\"33588886\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"YY1 transcription factor directly binds and transcriptionally activates the PFKP gene promoter in prostate cancer cells, forming an oncogenic YY1:BRD2/4-PFKP axis that sustains the Warburg effect; mutagenesis of YY1-bound cis-elements and gene loss-of-function/rescue studies confirmed the direct transcriptional regulation.\",\n      \"method\": \"Cistrome analysis (ChIP-seq), gene loss-of-function, YY1-bound cis-element mutagenesis, gene rescue studies, interactome profiling\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — cistrome mapping, mutagenesis of binding elements, epistasis rescue experiments, multiple orthogonal approaches\",\n      \"pmids\": [\"33849067\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PFKP is a nucleocytoplasmic shuttling protein containing functional nuclear export and nuclear localization sequences (NLS). Cyclin D3/CDK6 promotes PFKP nuclear translocation by dimerization and by exposing the NLS, enabling interaction with importin 9. In the nucleus, PFKP stimulates CXCR4 expression via c-Myc activity to promote T-ALL cell invasion.\",\n      \"method\": \"NLS/NES mapping, Cyclin D3/CDK6 overexpression, importin 9 co-immunoprecipitation, PFKP nuclear fractionation, CXCR4 expression assays, in vivo leukemia homing assays, CXCR4 antagonist rescue\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — nuclear localization signal mapping, protein-protein interaction (importin 9 Co-IP), c-Myc dependence, in vivo antagonist rescue, multiple orthogonal methods\",\n      \"pmids\": [\"34255748\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PFKP phosphorylates ATG4B at serine 34 (S34) in vitro, functioning as a protein kinase; amino acid deprivation strengthens the PFKP-ATG4B interaction, and this PFKP-mediated ATG4B phosphorylation enhances ATG4B activity and autophagic flux. PFKP S386 phosphorylation (under starvation) is required for ATG4B S34 phosphorylation.\",\n      \"method\": \"Tandem affinity purification-mass spectrometry, immunoprecipitation, in vitro kinase assay, CRISPR/Cas9 PFKP knockout, phosphosite mass spectrometry, autophagic flux assays\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro kinase assay directly demonstrating PFKP phosphorylates ATG4B-S34, confirmed by MS phosphosite mapping and CRISPR KO functional readout\",\n      \"pmids\": [\"33607258\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Mutant IDH1 upregulates PFKP expression through alteration of histone modification (as revealed by ChIP), and PFKP knockdown alleviates the mutant IDH1-induced increase in intrahepatic biliary organoid formation; high PFKP expression is more frequent in IDH-mutant cholangiocarcinoma.\",\n      \"method\": \"Gene expression analysis, ChIP for histone modification, Pfkp knockdown in intrahepatic biliary organoids, patient tissue immunohistochemistry\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP for epigenetic mechanism, organoid rescue experiment, single lab\",\n      \"pmids\": [\"31827136\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"PFKP interacts with AMPK; upon glucose starvation this interaction is greatly enhanced and promotes mitochondrial recruitment of AMPK, which then phosphorylates ACC2 to enhance long-chain fatty acid oxidation, maintaining energy/redox homeostasis and promoting cancer cell survival.\",\n      \"method\": \"Proteomics screening of AMPK-interacting proteins, co-immunoprecipitation, PFKP knockdown/overexpression, ACC2 phosphorylation assays, fatty acid oxidation assays, metabolic measurements\",\n      \"journal\": \"Cell discovery\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — proteomics-identified interaction validated by Co-IP, AMPK mitochondrial recruitment demonstrated, ACC2 phosphorylation as downstream readout, multiple orthogonal methods\",\n      \"pmids\": [\"35641476\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Fbxo7 promotes Cdk6-dependent phosphorylation of PFKP and Cdk6-independent ubiquitination of PFKP; loss of Fbxo7 reduces Cdk6 activity leading to increased glycolysis in CD4+ T cells, establishing PFKP as an essential Cdk6 substrate in T cells.\",\n      \"method\": \"Fbxo7 substrate screen, Fbxo7-deficient cells, metabolomics of activated CD4+ T cells, Cdk6 activity assays, ubiquitination assays, PFKP phosphorylation assays\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — substrate screen, Cdk6-dependent phosphorylation of PFKP validated, metabolomics confirmation, multiple orthogonal methods\",\n      \"pmids\": [\"35670764\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"PFKP promotes accumulation of its product fructose-1,6-bisphosphate (FBP); FBP inhibits RhoA/ROCK1 pathway-mediated cytoskeletal remodeling in podocytes, and PFKP overexpression rescues podocytes from high-glucose-induced cytoskeletal remodeling via this FBP-dependent mechanism.\",\n      \"method\": \"AAV-mediated PFKP overexpression/knockdown in mouse kidneys, siRNA in PTECs, targeted metabolomics, FBP measurement, FBP exogenous addition, RhoA/ROCK1 pathway assays, aldolase B manipulation\",\n      \"journal\": \"Frontiers in endocrinology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — metabolomics and direct FBP addition experiments, RhoA/ROCK1 pathway readout, in vivo AAV model, single lab\",\n      \"pmids\": [\"35095764\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"PFKP is lactylated at lysine 688 (K688), and this modification directly attenuates PFKP enzymatic activity, suggesting a negative feedback loop in glycolysis where lactate production leads to PFKP inhibition via lactylation.\",\n      \"method\": \"Mass spectrometry proteome-wide lactylation profiling, identification of PFKP K688 lactylation site, enzymatic activity assay of lactylated vs. non-lactylated PFKP\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — MS identification of modification site and in vitro enzymatic activity assay demonstrating inhibition, single lab\",\n      \"pmids\": [\"38155775\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CMBL (carboxymethylenebutenolidase-like) bridges TRIM25 E3 ubiquitin ligase to PFKP; ectopic CMBL enhances TRIM25 binding to PFKP, leading to PFKP ubiquitination and proteasomal degradation. p53 transcriptionally activates CMBL in response to genotoxic stress, repressing glycolysis via PFKP degradation.\",\n      \"method\": \"Co-immunoprecipitation (CMBL-TRIM25-PFKP ternary complex), ubiquitination assay, p53 transcriptional activation analysis, proteasome inhibition, CMBL/TRIM25 KO with PFKP rescue\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP of ternary complex, ubiquitination assay, genetic epistasis (p53→CMBL→TRIM25→PFKP), multiple orthogonal approaches\",\n      \"pmids\": [\"37967006\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"TGF-β1 recruits the SMAD3-SP1 transcriptional complex to the PFKP promoter (confirmed by ChIP-qPCR) to enhance PFKP expression, upregulating glycolysis in renal proximal tubular epithelial cells and promoting kidney fibrosis; PFKP knockdown or overexpression in AAV-transduced mice correspondingly reduced or promoted fibrosis.\",\n      \"method\": \"ChIP-qPCR, AAV-mediated PFKP overexpression/knockdown in mouse kidneys, TGF-β1 stimulation of PTECs, glycolysis assays, fibrosis histology\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ChIP-qPCR establishes direct SMAD3-SP1 binding to PFKP promoter, AAV in vivo functional validation, multiple orthogonal readouts\",\n      \"pmids\": [\"38086793\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"SIRT2 deacetylates PFKP at mouse lysine 394 (human K395), impairing its glycolytic function and reducing ATG4B phosphorylation; reduced ATG4B activation leads to decreased LC3 activation, suppressed LC3-associated phagocytosis (LAP), and impaired pathogen clearance in ethanol-exposed macrophages.\",\n      \"method\": \"Bone marrow-derived macrophages from SIRT2-deficient mice, SIRT2 pharmacological inhibition, PFKP acetylation/deacetylation assays, ATG4B phosphorylation, LC3 activation assays, phagocytosis/LAP assays, bacterial clearance and sepsis survival\",\n      \"journal\": \"Frontiers in immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — specific modification site identified (K394/K395), genetic and pharmacological SIRT2 inhibition, multiple downstream functional readouts, in vivo survival data\",\n      \"pmids\": [\"36865524\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Glycolytic enzyme PFKP acts as a protein kinase for Lin41 in mouse embryonic stem cells: Pfkp phosphorylates Lin41 at serine residues, stabilizing Lin41 by impeding its autoubiquitination and proteasomal degradation; this allows Lin41 to destabilize ectodermal specification mRNAs and favor endodermal differentiation.\",\n      \"method\": \"Pfkp genetic manipulation (Stat3 repression model), in vitro/in vivo kinase assay for Lin41 phosphorylation, Lin41 ubiquitination assays, mESC differentiation lineage marker assays\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro kinase assay establishing PFKP as kinase for Lin41, ubiquitination/stability assays, lineage differentiation functional outcome, multiple orthogonal methods\",\n      \"pmids\": [\"36660859\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Triptolide (TP) inhibits PFKP expression in Sertoli cells through the GATA4/Sp1 transcriptional axis: GATA4 regulates Sp1, which drives PFKP expression; suppression of GATA4 or Sp1 reduces PFKP levels and impairs glycolysis.\",\n      \"method\": \"GATA4/Sp1 knockdown, Sp1 pharmacological inhibitor (plicamycin), PFKP mRNA/protein assays, glucose/lactate assays, in vivo mouse TP dosing\",\n      \"journal\": \"Toxicology and applied pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic and pharmacological perturbation of transcriptional hierarchy, in vivo confirmation, single lab\",\n      \"pmids\": [\"34087332\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"PFKP confers cisplatin resistance in NSCLC by upregulating the drug efflux transporter ABCC2 through activation of the NF-κB pathway (increased phospho-IκBα and nuclear p65); PFKP knockdown reduces NF-κB activation and ABCC2 expression, sensitizing cells to cisplatin.\",\n      \"method\": \"PFKP overexpression/knockdown in NSCLC cell lines, luciferase assay for ABCC2 promoter, NF-κB signaling assays (p-IκBα, nuclear p65), cisplatin sensitivity (flow cytometry), in vivo xenograft\",\n      \"journal\": \"Translational lung cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — luciferase assay linking PFKP to ABCC2 promoter via NF-κB, multiple functional assays, in vivo validation, single lab\",\n      \"pmids\": [\"38090515\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"USP5 deubiquitinase directly binds PFKP and removes ubiquitin chains, stabilizing PFKP protein; USP5-mediated PFKP deubiquitination is essential for aerobic glycolysis and TNBC progression, with strong positive correlation between USP5 and PFKP levels in patient tissue.\",\n      \"method\": \"Co-immunoprecipitation, mass spectrometry protein identification, in vitro binding assay, ubiquitin assay, glycolysis assays, xenograft experiments, CPTAC database and IHC\",\n      \"journal\": \"Breast cancer research : BCR\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, in vitro binding assay, ubiquitination assay demonstrating deubiquitination, in vivo xenograft, MS identification, multiple orthogonal methods\",\n      \"pmids\": [\"38217030\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PFKP increases ERK-mediated stabilization of c-Myc protein, and c-Myc transcriptionally activates PFKP, forming a positive feedback loop that drives HNSCC progression; genetic and pharmacological co-targeting of PFKP and c-Myc shows synergistic anti-tumor effects in patient-derived organoids and xenografts.\",\n      \"method\": \"PFKP and c-Myc overexpression/knockdown, ERK signaling assays, c-Myc protein stability assays, c-Myc ChIP/transcriptional activation at PFKP, PDO and CDX/PDX models\",\n      \"journal\": \"Molecular cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis loop established by genetic perturbation and ChIP, multiple in vivo models, single lab\",\n      \"pmids\": [\"38982480\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF123 E3 ubiquitin ligase directly interacts with PFKP and induces its ubiquitination and degradation, thereby inhibiting glycolysis, cell cycle progression, and viability of breast cancer cells; overexpression of PFKP reverses the tumor-suppressive effects of RNF123.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination analysis, glycolysis assays, cell viability/cycle/colony formation assays, in vivo xenograft, lentiviral overexpression/silencing\",\n      \"journal\": \"Naunyn-Schmiedeberg's archives of pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP and ubiquitination assay, PFKP rescue experiment, single lab\",\n      \"pmids\": [\"39725718\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PFKP is lactylated at lysine 392 (K392); mutation of K392 diminishes PFKP lactylation and reduces glycolysis; PFKP K392 lactylation promotes glycolysis by regulating PTEN expression in hypoxia-treated ovarian cancer cells.\",\n      \"method\": \"Immunoprecipitation-western blot for PFKP lactylation, K392 site-directed mutagenesis, PFKP knockdown/overexpression, glycolysis assays, PTEN expression analysis, xenograft model\",\n      \"journal\": \"Biochemical genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — specific lactylation site mutagenesis, functional downstream PTEN regulation established, single lab\",\n      \"pmids\": [\"39638933\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PFKP binds AXL receptor tyrosine kinase and promotes its phosphorylation at Y779, activating AXL signaling and downstream MET phosphorylation in NSCLC cells, representing a non-metabolic oncogenic function of PFKP.\",\n      \"method\": \"PFKP knockdown (nanoparticle-mediated), co-immunoprecipitation of PFKP-AXL, AXL Y779 phosphorylation assays, MET phosphorylation assays, in vivo tumor growth\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP of PFKP-AXL interaction, phosphorylation readout at defined site, in vivo validation, single lab\",\n      \"pmids\": [\"39664584\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PFKP interacts with EIF2S2 (eukaryotic translation initiation factor 2 subunit beta); PFKP overexpression promotes new protein synthesis via EIF2S2, contributing to pathological cardiac hypertrophy; knockdown of EIF2S2 after PFKP overexpression reduces new protein synthesis and alleviates hypertrophy.\",\n      \"method\": \"Immunoprecipitation combined with LC-MS/MS to identify PFKP interactors, EIF2S2 knockdown rescue in NRCM and TAC mouse model, protein synthesis assays, cardiac phenotype characterization\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular basis of disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MS-identified interaction, genetic rescue epistasis (EIF2S2 KD reverses PFKP overexpression phenotype), in vivo TAC model, single lab\",\n      \"pmids\": [\"39419453\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PKP1 (Plakophilin-1) stabilizes PFKP by binding TRIM21 and preventing TRIM21-mediated ubiquitination and proteasomal degradation of PFKP; PKP1 depletion selectively reduces PFKP levels, and functional rescue with PFKP restores the proliferative phenotype driven by PKP1.\",\n      \"method\": \"CRISPR knockout screening, PKP1 depletion, PFKP ubiquitination assays, TRIM21 binding assay, PFKP rescue experiments, metabolic assays (OCR/ECAR), multiple LUSC cell lines\",\n      \"journal\": \"Biomarker research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ubiquitination assay, TRIM21 binding, functional rescue, multiple cell lines, single lab\",\n      \"pmids\": [\"40890861\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"HIF-1α transcriptionally controls PFKP mRNA in macrophages, as demonstrated by chromatin immunoprecipitation-qPCR (ChIP-qPCR); ethanol-induced oxidative stress impairs HIF-1α function, leading to reduced PFKP transcription, decreased glycolysis, and impaired phagocytosis; mitoquinol (MitoQ) restores HIF-1α function and PFKP expression.\",\n      \"method\": \"ChIP-qPCR for HIF-1α binding to PFKP promoter, macrophage ethanol exposure, PFKP mRNA/protein measurement, glycolysis assays, phagocytosis assays, MitoQ treatment, in vivo sepsis survival\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ChIP-qPCR directly demonstrates HIF-1α binding to PFKP promoter, pharmacological and genetic restoration of function, in vivo validation, orthogonal methods\",\n      \"pmids\": [\"40356076\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TRIS(2-chloroethyl)phosphate (TCEP) decreases PFKP glycolytic activity by enhancing O-linked N-acetylglucosamine (O-GlcNAc) transferase interaction with PFKP, thereby impairing glycolysis and ATP production in platelets, leading to suppression of platelet aggregation and activation.\",\n      \"method\": \"Proteomic analysis of platelets, O-GlcNAc transferase-PFKP interaction assay, PFKP activity assay, glycolysis/PPP pathway assays, ATP measurement, platelet activation assays, in vivo model\",\n      \"journal\": \"Environmental pollution\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — specific molecular interaction (OGT-PFKP) with PFKP activity readout, proteomics + functional assays, single lab\",\n      \"pmids\": [\"39828204\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Stat1 transcriptionally regulates Pfkp expression in macrophages; dsHMGB1 activates the Jak2/Stat1 pathway, which upregulates Pfkp to promote glycolysis and M1 macrophage polarization. ChIP-qPCR and dual-luciferase assays confirmed Stat1 binding to the Pfkp promoter.\",\n      \"method\": \"ChIP-qPCR, dual-luciferase assay, Jak2/Stat1 pathway inhibitor (fludarabine), dsHMGB1 treatment, glycolysis assays, macrophage polarization assays\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP-qPCR and luciferase confirm Stat1 binding to Pfkp promoter, pharmacological rescue, single lab\",\n      \"pmids\": [\"41079919\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"ATM kinase phosphorylates PFKP at threonine 278 (T278) in response to ionizing radiation and high glucose, promoting PFKP transition from tetramers to dimers; nuclear dimeric PFKP recruits casein kinase 2 (CK2), which phosphorylates RAD51 at T13 to enhance RAD51-BRCA2 interaction and homologous recombination (HR) repair.\",\n      \"method\": \"In vitro phosphorylation assay (ATM kinase), phosphosite mutagenesis (T278), PFKP tetramer/dimer structural analysis, nuclear fractionation, CK2 recruitment Co-IP, RAD51 T13 phosphorylation assay, HR repair assay, irradiation and high glucose conditions\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro kinase assay, specific phosphosite mutagenesis, structural transition (tetramer→dimer) demonstrated, downstream CK2-RAD51-BRCA2 cascade validated by Co-IP and phosphorylation assay, multiple orthogonal methods\",\n      \"pmids\": [\"42011782\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"PFKP directly binds AMOTL1 and inhibits its ubiquitin-mediated degradation; PFKP-driven aerobic glycolysis and EMT in HNSCC are AMOTL1-dependent. PFKP promotes YAP nuclear translocation via AMOTL1, suppressing Hippo pathway activity and amplifying glycolytic flux.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination analysis, AMOTL1 knockdown rescue, YAP nuclear translocation assays, Hippo pathway readouts, in vivo nude mice tumor models\",\n      \"journal\": \"Journal of translational internal medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP, ubiquitination assay, AMOTL1 genetic rescue, in vivo confirmation, single lab\",\n      \"pmids\": [\"41727965\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"USP14 deubiquitinase stabilizes PFKP through K48-linked deubiquitination; c-Myc transcriptionally upregulates USP14, while PFKP enhances ERK-dependent c-Myc protein stability, forming a c-Myc-USP14-PFKP feed-forward regulatory circuit that sustains PDAC chemoresistance.\",\n      \"method\": \"Ubiquitination assays (K48-linked), USP14-PFKP protein interaction studies, c-Myc transcriptional activation of USP14, ERK-c-Myc stability assays, patient-derived organoids, syngeneic and xenograft models, combined targeting experiments\",\n      \"journal\": \"Drug resistance updates\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — K48-specific ubiquitination assay, feed-forward circuit validated by multiple genetic/biochemical approaches, in vivo models, single lab\",\n      \"pmids\": [\"41812332\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"PFKP K688 lactylation (K688la) is induced by hypoxia/ischemia and enhances PFKP enzymatic activity (distinct from K688 lactylation inhibiting activity in cancer cells [PMID 38155775]); a K688E mutation mimicking hyper-lactylation further amplifies glycolysis and confers cardiomyocyte cytoprotection.\",\n      \"method\": \"Lactylome proteomics (LAD-operated mice, hypoxic cardiomyocytes), PFKP K688E mutagenesis, PFKP enzymatic activity assays, ECAR/OCR metabolic assays, cardiomyocyte survival assays, 2-DG rescue\",\n      \"journal\": \"Frontiers in pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — site-directed mutagenesis at K688 with enzymatic activity readout, in vivo and in vitro lactylome proteomics, single lab; note: contradicts PMID 38155775 on K688 lactylation effect direction\",\n      \"pmids\": [\"41919225\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"PFKP (phosphofructokinase, platelet isoform) is a rate-limiting glycolytic enzyme that catalyzes the phosphorylation of fructose-6-phosphate to fructose-1,6-bisphosphate, but it also functions as a non-canonical protein kinase (phosphorylating ATG4B-S34 to regulate autophagy and Lin41-serine to direct stem cell differentiation), undergoes nucleocytoplasmic shuttling (driven by Cyclin D3/CDK6-importin 9 interaction) to transcriptionally activate CXCR4 via c-Myc, and upon ATM-mediated T278 phosphorylation recruits CK2 to phosphorylate RAD51-T13 for homologous recombination repair; its activity and stability are regulated by multiple post-translational modifications including ubiquitination by HRD1, TRIM25/CMBL, RNF123, and USP14 (stabilized by USP5 and PKP1/TRIM21 competition), phosphorylation by CDK6 (via Fbxo7), deacetylation by SIRT2 at K394/K395 (reducing activity and LAP-mediated phagocytosis), and lactylation at K688 (with context-dependent activity effects); its transcription is activated by KLF4, YY1:BRD2/4, HIF-1α, Stat1, SMAD3-SP1, and c-Myc, and repressed by Snail, Stat3, and p53/CMBL, collectively placing PFKP at the intersection of glycolytic flux control, immune function, autophagy, DNA damage repair, and oncogenic signaling.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"PFKP (platelet-type phosphofructokinase) is a rate-limiting glycolytic enzyme whose expression, stability, and moonlighting activities place it at the convergence of metabolic flux control, autophagy, DNA repair, and oncogenic signaling [#2, #9, #31]. Its glycolytic output is controlled at multiple levels: a broad transcriptional network activates PFKP — KLF4 [#1], YY1 with BRD2/4 [#7], HIF-1\\u03b1 [#28], Stat1 [#30], SMAD3\\u2013SP1 [#16], GATA4/Sp1 [#19], and a c-Myc positive-feedback loop [#22] — while Snail represses it to divert glucose into the pentose phosphate pathway [#2], and p53 represses it indirectly by inducing CMBL [#15]. PFKP protein stability is set by competing ubiquitin machineries, with the E3 ligases HRD1 [#6], TRIM25 (bridged by CMBL) [#15], RNF123 [#23], and TRIM21 [#27] driving proteasomal degradation, and the deubiquitinases USP5 [#21] and USP14 [#33] (the latter in a c-Myc\\u2013USP14\\u2013PFKP feed-forward circuit) stabilizing it. PFKP enzymatic activity is further tuned by post-translational modification, including SIRT2-mediated deacetylation at K394/K395 [#17], lactylation at K688 and K392 [#14, #24, #34], and O-GlcNAcylation [#29]. Beyond catalysis, PFKP acts as a non-canonical protein kinase, phosphorylating ATG4B-S34 to promote autophagic flux [#9] and Lin41 to stabilize it and direct endodermal differentiation in stem cells [#18]. PFKP shuttles between cytoplasm and nucleus via mapped NLS/NES sequences, with Cyclin D3/CDK6 and importin 9 driving nuclear entry to transcriptionally activate CXCR4 through c-Myc [#8]; upon ATM-mediated T278 phosphorylation it shifts from tetramer to dimer and recruits CK2 to phosphorylate RAD51-T13, supporting homologous recombination repair [#31]. It also exerts protein-binding functions independent of catalysis, recruiting AMPK to mitochondria to drive fatty acid oxidation [#11], activating AXL receptor signaling [#25], and stabilizing AMOTL1 to promote YAP nuclear translocation [#32].\",\n  \"teleology\": [\n    {\n      \"year\": 1983,\n      \"claim\": \"Establishing the chromosomal location of the platelet PFK subunit gene was the first step in defining PFKP as a distinct, dosage-sensitive genetic locus.\",\n      \"evidence\": \"Somatic cell hybrid panel with active-enzyme immunoprecipitation and gene-dosage analysis in fibroblasts\",\n      \"pmids\": [\"6222962\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not address tissue-specific regulation or non-glycolytic functions\", \"No protein-level mechanism beyond enzyme dosage\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Identifying KLF4 as a direct PFKP promoter activator showed PFKP transcription is selectively wired into oncogenic glycolytic programs distinct from other PFK isoforms.\",\n      \"evidence\": \"Promoter binding (ChIP/reporter), KLF4 knockdown/overexpression and glucose/lactate assays in breast cancer cells\",\n      \"pmids\": [\"21586797\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab\", \"Did not connect KLF4 to upstream signals controlling PFKP\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Demonstrating Snail-mediated PFKP repression established PFKP as a metabolic switch directing glucose between glycolysis and the pentose phosphate pathway during EMT.\",\n      \"evidence\": \"Snail and PFKP knockdown/rescue with metabolic flux assays and in vivo metastasis models\",\n      \"pmids\": [\"28176759\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of Snail recruitment to PFKP locus not detailed\", \"PPP rerouting quantified but enzymatic regulation downstream of repression not addressed\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Linking PFKP to VDAC2 at mitochondria revealed that spatial restriction of PFKP activity controls glioma stemness.\",\n      \"evidence\": \"Co-localization, VDAC2 manipulation, PFK inhibitor rescue and stem/tumorigenicity assays\",\n      \"pmids\": [\"30250190\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Interaction inferred from co-localization and inhibitor rescue rather than direct binding\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Defining an ATM\\u2192HIF1A\\u2192PFKP translational axis under hypoxia connected PFKP-derived citrate accumulation to invasive signaling.\",\n      \"evidence\": \"ATM inhibition/knockdown, HIF1A studies, citrate measurement and invasion/xenograft assays in breast cancer\",\n      \"pmids\": [\"30850587\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Translational regulation mechanism not molecularly resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Mutant IDH1 was shown to upregulate PFKP via altered histone modification, tying oncometabolite-driven epigenetics to glycolytic gene expression.\",\n      \"evidence\": \"Expression analysis, ChIP for histone marks, organoid PFKP knockdown and patient IHC\",\n      \"pmids\": [\"31827136\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Specific histone mark and reader at PFKP locus not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Multiple 2021 studies revealed PFKP is controlled post-transcriptionally (FTO/m6A/YTHDF2), by ubiquitin-mediated degradation (HRD1), and by direct transcriptional activation (YY1:BRD2/4), broadening the regulatory network governing PFKP abundance.\",\n      \"evidence\": \"m6A-seq with genetic rescue in leukemia; MS, reciprocal Co-IP and ubiquitination assays for HRD1; cistrome mapping and cis-element mutagenesis for YY1\",\n      \"pmids\": [\"33434505\", \"33588886\", \"33849067\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How these layers integrate to set net PFKP levels in a given cell not addressed\", \"Each established in distinct cancer contexts\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Discovery that PFKP phosphorylates ATG4B-S34 redefined PFKP as a non-canonical protein kinase coupling metabolic state to autophagy.\",\n      \"evidence\": \"TAP-MS, in vitro kinase assay, phosphosite MS and CRISPR knockout autophagic flux assays under amino acid deprivation\",\n      \"pmids\": [\"33607258\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of PFKP kinase activity unresolved\", \"Whether kinase and glycolytic activities are mutually exclusive not addressed\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Mapping functional NLS/NES sequences and Cyclin D3/CDK6\\u2013importin 9-driven nuclear import established PFKP as a shuttling protein with a nuclear transcriptional function (CXCR4 via c-Myc).\",\n      \"evidence\": \"NLS/NES mapping, importin 9 Co-IP, nuclear fractionation, CXCR4 assays and in vivo homing/antagonist rescue in T-ALL\",\n      \"pmids\": [\"34255748\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether nuclear PFKP retains catalytic activity not resolved\", \"Direct DNA contact vs c-Myc cofactor role not distinguished\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identifying PFKP\\u2013AMPK and CDK6\\u2013Fbxo7 axes showed PFKP both directs mitochondrial AMPK signaling under starvation and serves as an essential CDK6 substrate in T cells.\",\n      \"evidence\": \"Proteomics and Co-IP with ACC2 phosphorylation/FAO assays (AMPK); Fbxo7 substrate screen with metabolomics and ubiquitination/phosphorylation assays in CD4+ T cells (CDK6/Fbxo7)\",\n      \"pmids\": [\"35641476\", \"35670764\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional residue(s) of CDK6 phosphorylation on PFKP not mapped\", \"Crosstalk between PFKP scaffolding and catalysis not resolved\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Demonstrating that PFKP product FBP inhibits RhoA/ROCK1 in podocytes linked PFKP catalytic output to cytoskeletal protection in a non-cancer setting.\",\n      \"evidence\": \"AAV PFKP manipulation in mouse kidney, targeted metabolomics, exogenous FBP addition and RhoA/ROCK1 readouts\",\n      \"pmids\": [\"35095764\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct FBP\\u2013RhoA/ROCK1 mechanism not biochemically defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"A series of 2023 studies established the PTM and degradation logic of PFKP: lactylation (K688), deacetylation (K394/K395 by SIRT2), and CMBL-bridged TRIM25 ubiquitination under p53 control, each tuning enzymatic activity or stability.\",\n      \"evidence\": \"MS lactylation profiling with activity assays; SIRT2 KO/inhibition with ATG4B/LAP readouts; reciprocal Co-IP of CMBL\\u2013TRIM25\\u2013PFKP with ubiquitination and p53 epistasis\",\n      \"pmids\": [\"38155775\", \"36865524\", \"37967006\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Interplay among competing PTMs on the same protein not integrated\", \"Stoichiometry and dynamics of modifications in vivo unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identifying PFKP as a kinase for Lin41 extended its non-canonical kinase role to stem cell lineage control, stabilizing Lin41 to favor endodermal differentiation.\",\n      \"evidence\": \"Pfkp manipulation, in vitro/in vivo Lin41 kinase and ubiquitination assays, mESC lineage marker assays\",\n      \"pmids\": [\"36660859\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Lin41 target serine residues not fully enumerated\", \"Relationship to ATG4B kinase activity not addressed\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Adding SMAD3-SP1 (TGF-\\u03b21), GATA4/Sp1, and NF-\\u03baB\\u2013ABCC2 links expanded PFKP's roles to kidney fibrosis, Sertoli cell glycolysis, and NSCLC drug resistance.\",\n      \"evidence\": \"ChIP-qPCR with AAV in vivo fibrosis models; GATA4/Sp1 knockdown with glycolysis assays; PFKP\\u2013NF-\\u03baB\\u2013ABCC2 luciferase and cisplatin sensitivity assays\",\n      \"pmids\": [\"38086793\", \"34087332\", \"38090515\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Each axis from a single lab\", \"Whether glycolytic vs moonlighting activity drives these phenotypes not always distinguished\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"2024 work deepened the PFKP stability and signaling map, adding USP5 and RNF123 to the ubiquitin axis, a c-Myc/ERK feedback loop, and lactylation at K392 regulating PTEN.\",\n      \"evidence\": \"Co-IP, in vitro binding and ubiquitination assays (USP5, RNF123); ERK/c-Myc stability and ChIP (HNSCC); K392 mutagenesis with PTEN/glycolysis readouts (ovarian cancer)\",\n      \"pmids\": [\"38217030\", \"39725718\", \"38982480\", \"39638933\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Competing E3/DUB activities not reconciled into a unified stability model\", \"Most findings single-lab and context-specific\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identifying PFKP binding partners AXL, EIF2S2, and AMOTL1 expanded its catalysis-independent scaffolding functions into receptor signaling, protein synthesis, and cardiac hypertrophy.\",\n      \"evidence\": \"Co-IP/MS with AXL Y779 and MET phosphorylation assays; LC-MS/MS with EIF2S2 rescue in TAC model; AMOTL1 binding/ubiquitination and YAP/Hippo assays\",\n      \"pmids\": [\"39664584\", \"39419453\", \"41727965\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs bridged interactions not always distinguished\", \"Single-lab findings across distinct disease contexts\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Showing HIF-1\\u03b1, Stat1, and PKP1/TRIM21 each control PFKP refined macrophage and squamous-cancer regulation, including a PKP1-mediated block of TRIM21 ubiquitination.\",\n      \"evidence\": \"ChIP-qPCR and pharmacological rescue for HIF-1\\u03b1 and Stat1 in macrophages; CRISPR screening, TRIM21 binding and ubiquitination/rescue assays for PKP1 in LUSC\",\n      \"pmids\": [\"40356076\", \"41079919\", \"40890861\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"O-GlcNAc and other PTM cross-regulation in immune cells not integrated\", \"Most studies single-lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstrating that O-GlcNAc transferase interaction lowers PFKP activity in platelets connected PFKP-controlled glycolysis to platelet aggregation and a toxicant exposure phenotype.\",\n      \"evidence\": \"Platelet proteomics, OGT\\u2013PFKP interaction and activity assays with ATP/aggregation readouts\",\n      \"pmids\": [\"39828204\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"O-GlcNAc site on PFKP not mapped\", \"Single lab\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Defining ATM\\u2013T278 phosphorylation that drives a tetramer-to-dimer transition and nuclear CK2 recruitment established PFKP as a direct participant in homologous recombination DNA repair.\",\n      \"evidence\": \"In vitro ATM kinase assay, T278 mutagenesis, oligomeric state analysis, CK2 Co-IP and RAD51-T13/HR repair assays under irradiation and high glucose\",\n      \"pmids\": [\"42011782\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of dimer-specific CK2 recruitment not resolved\", \"Whether dimeric PFKP retains glycolytic catalysis unclear\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Adding a c-Myc\\u2013USP14\\u2013PFKP feed-forward circuit and a context-dependent K688 lactylation that enhances activity in cardiomyocytes refined PFKP stability control and reconciled tissue-specific PTM effects.\",\n      \"evidence\": \"K48-linked ubiquitination, USP14 interaction and ERK\\u2013c-Myc circuit in PDAC models; cardiac lactylome proteomics with K688E mutagenesis and activity/survival assays\",\n      \"pmids\": [\"41812332\", \"41919225\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Opposite directionality of K688 lactylation effect between cancer and cardiomyocytes mechanistically unexplained\", \"Single-lab findings\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How PFKP's catalytic, kinase, scaffolding, and nuclear functions are coordinately switched within a single cell, and what structural features distinguish its glycolytic from non-canonical activities, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model reconciling tetramer/dimer states with kinase and scaffold activities\", \"Integration of competing E3/DUB and PTM inputs into net activity unknown\", \"Context-dependent lactylation effects unexplained at the mechanistic level\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [9, 18, 31]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [9, 18]},\n      {\"term_id\": \"GO:0016301\", \"supporting_discovery_ids\": [9, 18, 31]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [6]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [8, 31]},\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [3, 11]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [2, 6, 21]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [9, 17]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [31]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [6, 15, 21, 33]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [17, 28, 30]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [25, 32]}\n    ],\n    \"complexes\": [\"CMBL\\u2013TRIM25\\u2013PFKP ubiquitination complex\"],\n    \"partners\": [\"ATG4B\", \"AMPK\", \"VDAC2\", \"AXL\", \"AMOTL1\", \"EIF2S2\", \"TRIM25\", \"USP5\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"tie","faith_supported":7,"faith_total":7,"faith_pct":100.0}}