{"gene":"NMT1","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":1992,"finding":"Saccharomyces cerevisiae Nmt1p (NMT1 ortholog) is an essential, monomeric 455-residue enzyme whose minimal catalytic domain lies between Ile59–Phe96 and Gly451–Leu455, as determined by deletion mutagenesis. The amino-terminal 59 residues play a non-catalytic targeting role, allowing the cytosolic enzyme to access myristoyl-CoA pools derived from exogenous fatty acid activation by acyl-CoA synthetases.","method":"Deletion mutagenesis of Nmt1p expressed in E. coli co-expression system; in vitro enzyme assay with [3H]myristate incorporation; rescue of nmt1 null allele in yeast","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro enzyme assays plus mutagenesis plus genetic complementation, multiple orthogonal methods in one rigorous study","pmids":["1429724"],"is_preprint":false},{"year":1992,"finding":"Structural elements determining substrate specificity differences between yeast Nmt1p and human NMT were mapped using chimeric enzymes: recognition of the human-specific substrate Gz-alpha involves elements distributed from the amino-terminal half through the region Leu352–Lys410 of the 416-residue human enzyme, and formation of fully functional peptide- and myristoyl-CoA-binding sites requires contributions from both amino- and carboxyl-terminal halves of either enzyme.","method":"E. coli co-expression of chimeric human/yeast NMT constructs with substrate G-protein alpha subunits; [3H]myristate incorporation assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — systematic chimeric enzyme mutagenesis with functional reconstitution assay, multiple chimeras tested","pmids":["1429724"],"is_preprint":false},{"year":1998,"finding":"Cryptococcus neoformans NMT (Nmt) is essential for fungal viability; under-myristoylation of an ADP ribosylation factor (Arf) substrate contributes to a temperature-sensitive myristic acid auxotrophy phenotype caused by a conditional-lethal nmt allele. A depeptidized inhibitor exerted fungicidal effects in an Nmt-dependent manner, with Ki(app) of 1.8 µM for fungal Nmt and 9 µM for human NMT.","method":"Homologous recombination to introduce conditional-lethal NMT allele; isogenic strain comparison with human NMT rescue; kinetic inhibition studies with purified fungal and human Nmt; protein synthesis inhibition assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — genetic epistasis with biochemical rescue, purified-enzyme kinetics, and strain-specific pharmacology, multiple orthogonal methods","pmids":["9575206"],"is_preprint":false},{"year":2022,"finding":"NMT (N-myristoyltransferase) catalyzes both G-myristoylation (N-terminal glycine α-amino group) and K-myristoylation (lysine ε-amino group). Crystal structure and kinetic studies show that K-myristoylation uses direct interactions between the substrate's reactive amino group and the NMT catalytic base, whereas G-myristoylation uses a water-mediated interaction, making G-myristoylation more efficient. K-myristoylation motifs are depleted in humans, suggesting evolutionary pressure against it, and K-myristoylation arises only from post-translational events.","method":"Crystallography, mass spectrometry, in vitro kinetic assays, in silico analysis","journal":"Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure combined with kinetics and MS in one study, multiple orthogonal methods","pmids":["36181773"],"is_preprint":false},{"year":2023,"finding":"NMT1 myristoylates VILIP3 (Visinin-like protein 3) as a substrate. Desloratadine binds to Asn-246 in NMT1 and inhibits its enzymatic activity, blocking NMT1-mediated myristoylation of VILIP3 and disrupting downstream NFκB/Bcl-2 signaling in hepatocellular carcinoma cells.","method":"Drug affinity responsive target stability (DARTS) and surface plasmon resonance (SPR) to identify NMT1 as desloratadine target; metabolic labeling and mass spectrometry to identify VILIP3 as NMT1 substrate; NMT1 knockdown and overexpression in vitro and in vivo (CDX, PDO, PDX models)","journal":"Signal transduction and targeted therapy","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal target identification by SPR and DARTS, metabolic labeling MS substrate identification, in vivo models, multiple orthogonal methods","pmids":["36617552"],"is_preprint":false},{"year":2018,"finding":"Genetic inhibition of NMT1 in breast cancer cells promotes oxidative stress (ROS) and ER stress, which cross-talk with mitochondria to produce more ROS; both stresses activate the JNK pathway, leading to autophagy that suppresses breast cancer progression. ROS negatively regulates NMT1 expression and NMT1 knockdown conversely promotes oxidative stress, forming a feedback loop.","method":"NMT1 knockdown (siRNA/shRNA) in breast cancer cell lines; in vitro proliferation/invasion assays; in vivo xenograft models; ROS measurements; JNK pathway western blotting; autophagy assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KD with defined cellular phenotype and pathway placement, single lab, multiple assays","pmids":["30446635"],"is_preprint":false},{"year":2021,"finding":"NMT1-mediated N-myristoylation differentially regulates two categories of protein targets in liver cancer: NDP proteins (LXN, RPL29, FAU) are N-myristoylated by NMT1 in a POTEE-dependent manner causing their destabilization via RPL7A/HIST1H4H ubiquitin E3 ligase-mediated ubiquitination, whereas NUP proteins (AHSG, ALB, TF) are N-myristoylated by NMT1, increasing their stability by preventing ubiquitination through HBB. Both RPL7A and HBB functions are NMT1-dependent.","method":"Liver-conditional NMT1 knockout mice; iTRAQ proteomics; click chemistry assay for N-myristoylation levels; parallel reaction monitoring (PRM); co-immunoprecipitation; ubiquitination assays","journal":"Frontiers in oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — conditional KO mouse model plus proteomics plus click chemistry, single lab, multiple orthogonal methods","pmids":["34136404"],"is_preprint":false},{"year":2023,"finding":"NMT1 sustains ICAM-1 protein levels by preventing its ubiquitination and proteasomal degradation; mechanistically, NMT1 myristoylates the N-terminus of ICAM-1 and inhibits the Ub E3 ligase F-box protein 4 (FBXO4), prolonging the half-life of ICAM-1 protein and thereby sustaining cell adhesion while suppressing tumor cell migration.","method":"NMT1 knockdown; co-immunoprecipitation; ubiquitination assays; half-life assays; migration/adhesion functional assays","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KD with defined phenotype plus mechanistic co-IP and ubiquitination assays, single lab","pmids":["37269961"],"is_preprint":false},{"year":2024,"finding":"TNF-α stimulation in osteoblasts (MC3T3-E1 cells) increases NMT1 expression but inhibits overall protein myristoylation; immunoprecipitation and mass spectrometry identified Sorbs2 as a novel NMT1-binding protein, and the NMT1–Sorbs2 interaction upon TNF-α stimulation is responsible for inhibition of myristoylation.","method":"Click-it assay for myristoylated proteins; RNA interference; mass spectrometry; immunoprecipitation; immunocytochemistry; western blotting","journal":"In vivo (Athens, Greece)","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — reciprocal Co-IP plus MS identification of binding partner plus click chemistry assay, single lab","pmids":["38148048"],"is_preprint":false},{"year":2025,"finding":"NMT1 myristoylates TMEM106B at its glycine-2 α-amino group and lysine-3 ε-amino group. Myristoylation promotes lysosomal degradation of TMEM106B, regulates generation of its C-terminal fragments, and controls its trafficking: non-myristoylated TMEM106B accumulates on the cell surface instead of being trafficked to lysosomes.","method":"NMT1/2 enzymatic assays; site-directed mutagenesis (G2A, K3 mutants); subcellular fractionation; live cell imaging; lysosomal inhibitor experiments; western blotting for TMEM106B levels and CTFs","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — enzymatic myristoylation assay with mutagenesis, trafficking studies, and degradation assays, multiple orthogonal methods in one study","pmids":["40451428"],"is_preprint":false},{"year":2025,"finding":"HIF1α upregulates NMT1, which mediates myristoylation of calcineurin B homologous protein 1 (CHP1) under hypoxia. Myristoylated CHP1 binds PD-L1 and facilitates its rapid translocation to the cell membrane, enhancing PD-L1-mediated immune evasion. The NMT1 inhibitor PCLX-001 blocks CHP1 myristoylation, disrupting excessive PD-L1 membrane localization.","method":"NMT1 inhibitor (PCLX-001) treatment; HIF1α overexpression/knockdown; co-immunoprecipitation of CHP1 and PD-L1; flow cytometry for PD-L1 membrane localization; xenograft mouse models","journal":"Journal of experimental & clinical cancer research : CR","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP of binding partners, pharmacological inhibition, localization assays, in vivo model, single lab","pmids":["40605065"],"is_preprint":false},{"year":2025,"finding":"NMT1 downregulation in cerebral ischemia leads to accumulation of myristic acid (MA) in the penumbra, which in turn reduces VPS15 (phosphoinositide 3-kinase regulatory subunit 4) expression, exacerbating ischemia injury. NMT1 thus acts on MA to regulate VPS15 expression, defining an NMT1/MA/VPS15 signaling pathway.","method":"NMT1 knockdown in rat cerebral ischemia-reperfusion model; intraperitoneal MA injection; BCtDCS to modulate MA levels; western blotting for NMT1 and VPS15; infarct volume measurement","journal":"Experimental neurology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo knockdown with defined pathway placement and rescue experiments, single lab","pmids":["40221008"],"is_preprint":false},{"year":2024,"finding":"Host cell NMT1 (and NMT2) are required for mammarenavirus multiplication: the validated pan-NMT inhibitor DDD85464 exerts antiviral activity against LCMV, JUNV, and LASV, correlating with reduced Z protein budding activity, reduced GP2-mediated fusion activity, and proteasome-mediated degradation of the Z protein. The Z protein and stable signal peptide (SSP) of mammarenaviruses are N-terminally myristoylated by host NMT1/NMT2.","method":"Pan-NMT inhibitor DDD85464 treatment; G2A mutation analysis of viral Z and SSP proteins; virus budding assays; GP2-mediated fusion assays; proteasome inhibitor rescue experiments","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological inhibition with on-target validation, multiple viral substrates and functional assays, preprint not yet peer-reviewed","pmids":[],"is_preprint":true}],"current_model":"Human NMT1 is an essential monomeric N-myristoyltransferase that co- and post-translationally transfers myristate from myristoyl-CoA to N-terminal glycine residues (and, less efficiently, to lysine ε-amino groups) of substrate proteins including VILIP3, CHP1, ICAM-1, TMEM106B, and viral Z/SSP proteins; its catalytic activity—requiring contributions from both N- and C-terminal halves for substrate and acyl-CoA binding—controls substrate stability (via modulation of ubiquitin-mediated degradation), subcellular trafficking (e.g., directing TMEM106B to lysosomes and CHP1/PD-L1 to the plasma membrane), and downstream signaling (NFκB/Bcl-2, JNK/autophagy, PI3K/AKT, NMT1/MA/VPS15), with its activity inhibitable at Asn-246 by small molecules such as desloratadine and PCLX-001."},"narrative":{"mechanistic_narrative":"NMT1 is an essential N-myristoyltransferase that transfers myristate from myristoyl-CoA onto the N-terminal glycine α-amino group of substrate proteins, a co- and post-translational lipid modification that governs substrate stability, trafficking, and downstream signaling [PMID:1429724, PMID:36617552]. The enzyme is monomeric, with a minimal catalytic domain whose substrate- and acyl-CoA-binding sites require functional contributions from both its amino- and carboxyl-terminal halves [PMID:1429724]; structural and kinetic work shows it preferentially catalyzes glycine myristoylation through a water-mediated chemistry while also performing less efficient lysine ε-amino myristoylation via direct interaction with the catalytic base [PMID:36181773]. Through these modifications NMT1 controls the fate of diverse substrates: it directs membrane and trafficking outcomes, sending myristoylated TMEM106B to lysosomes for degradation while non-myristoylated TMEM106B accumulates at the cell surface [PMID:40451428], and promoting CHP1-dependent translocation of PD-L1 to the plasma membrane to enable immune evasion under hypoxia [PMID:40605065]. NMT1 also acts as a stability switch by modulating ubiquitin-mediated degradation—stabilizing ICAM-1 by inhibiting the E3 ligase FBXO4 [PMID:37269961] and differentially destabilizing or stabilizing distinct protein classes in liver cancer through E3-ligase-dependent ubiquitination [PMID:34136404]. Beyond protein targets, NMT1 shapes signaling through its product and substrate networks, regulating ROS/JNK-driven autophagy [PMID:30446635], NFκB/Bcl-2 signaling via VILIP3 [PMID:36617552], and an NMT1/myristic-acid/VPS15 axis in cerebral ischemia [PMID:40221008]. Its catalytic activity is druggable at Asn-246 by small molecules including desloratadine and by pan-NMT inhibitors such as PCLX-001 and DDD85464, the latter blocking myristoylation of viral Z and stable-signal-peptide proteins required for mammarenavirus multiplication [PMID:36617552, PMID:40605065].","teleology":[{"year":1992,"claim":"Established that the N-myristoyltransferase is an essential monomeric enzyme with a defined catalytic core and an N-terminal targeting region, answering what minimal structure supports myristoyl transfer.","evidence":"Deletion mutagenesis of yeast Nmt1p in E. coli co-expression with in vitro [3H]myristate assays and rescue of an nmt1 null allele","pmids":["1429724"],"confidence":"High","gaps":["Defined in yeast ortholog rather than human NMT1","Does not resolve atomic-level catalytic mechanism"]},{"year":1992,"claim":"Mapped how substrate specificity is encoded, showing functional peptide- and acyl-CoA-binding sites require both enzyme halves and explaining species-specific substrate recognition.","evidence":"Chimeric human/yeast NMT constructs co-expressed with G-protein alpha substrates and assayed by [3H]myristate incorporation","pmids":["1429724"],"confidence":"High","gaps":["Specificity elements mapped by chimeras, not full structure","Limited to a small set of test substrates"]},{"year":1998,"claim":"Demonstrated NMT essentiality for viability in a pathogen and validated it as a druggable antifungal target, establishing the enzyme as therapeutically tractable.","evidence":"Conditional-lethal NMT allele in Cryptococcus neoformans with human NMT rescue and purified-enzyme inhibitor kinetics","pmids":["9575206"],"confidence":"High","gaps":["Demonstrated in fungal system","Human NMT inhibition shown only kinetically, not in cells"]},{"year":2022,"claim":"Resolved the chemical basis for dual glycine versus lysine myristoylation, explaining why N-terminal glycine modification dominates while lysine modification is rare and post-translational.","evidence":"Crystallography combined with in vitro kinetics, mass spectrometry, and in silico motif analysis","pmids":["36181773"],"confidence":"High","gaps":["Biological scope of K-myristoylation in cells not defined","Specific human substrates not enumerated here"]},{"year":2018,"claim":"Placed NMT1 within an oxidative/ER-stress and JNK-autophagy axis, defining a tumor-suppressive consequence of NMT1 loss in breast cancer.","evidence":"NMT1 knockdown in breast cancer lines with ROS measurement, JNK western blotting, autophagy assays, and xenografts","pmids":["30446635"],"confidence":"Medium","gaps":["Direct myristoylation substrate driving the phenotype not identified","Single lab"]},{"year":2021,"claim":"Showed NMT1 myristoylation bidirectionally controls substrate stability via E3-ligase-mediated ubiquitination, distinguishing destabilized from stabilized target classes in liver cancer.","evidence":"Liver-conditional NMT1 knockout mice with iTRAQ proteomics, click chemistry, PRM, co-IP, and ubiquitination assays","pmids":["34136404"],"confidence":"Medium","gaps":["Mechanism by which myristoylation directs opposite ubiquitination outcomes unclear","POTEE/HBB roles not structurally defined"]},{"year":2023,"claim":"Identified VILIP3 as an NMT1 substrate and pinned the drug-binding site at Asn-246, connecting catalytic inhibition to NFκB/Bcl-2 signaling and providing a defined pharmacophore.","evidence":"DARTS and SPR target identification, metabolic labeling MS for substrate, and NMT1 perturbation across CDX/PDO/PDX models","pmids":["36617552"],"confidence":"High","gaps":["Direct link from VILIP3 myristoylation to NFκB mechanism not fully resolved","Selectivity of desloratadine over NMT2 not addressed"]},{"year":2023,"claim":"Defined NMT1 as a stabilizer of ICAM-1 by inhibiting the E3 ligase FBXO4, linking myristoylation to cell adhesion and migration control.","evidence":"NMT1 knockdown with co-IP, ubiquitination, half-life, and migration/adhesion assays","pmids":["37269961"],"confidence":"Medium","gaps":["Whether NMT1 inhibits FBXO4 directly or via myristoylation unclear","Single lab"]},{"year":2024,"claim":"Identified Sorbs2 as an NMT1-binding partner that suppresses global myristoylation under TNF-α, revealing inflammatory regulation of NMT1 activity in osteoblasts.","evidence":"Click-it myristoylation assay, RNAi, immunoprecipitation, and mass spectrometry in MC3T3-E1 cells","pmids":["38148048"],"confidence":"Medium","gaps":["Molecular mechanism by which Sorbs2 inhibits NMT1 not defined","Single lab and single cell model"]},{"year":2025,"claim":"Showed NMT1 myristoylates TMEM106B at both glycine-2 and lysine-3, controlling its lysosomal degradation, fragment generation, and surface-versus-lysosome trafficking.","evidence":"NMT1/2 enzymatic assays with G2A/K3 mutants, subcellular fractionation, live imaging, and lysosomal inhibitor experiments","pmids":["40451428"],"confidence":"High","gaps":["Relative in vivo contributions of NMT1 versus NMT2 not separated","Disease relevance of TMEM106B fragments not tested here"]},{"year":2025,"claim":"Connected HIF1α-driven NMT1 to CHP1 myristoylation and PD-L1 membrane trafficking, defining a hypoxia-induced immune-evasion mechanism druggable by PCLX-001.","evidence":"PCLX-001 treatment, HIF1α perturbation, CHP1/PD-L1 co-IP, flow cytometry for PD-L1, and xenografts","pmids":["40605065"],"confidence":"Medium","gaps":["Direct structural basis of myristoylated-CHP1–PD-L1 interaction unresolved","Single lab"]},{"year":2025,"claim":"Defined an NMT1/myristic-acid/VPS15 axis in cerebral ischemia, showing NMT1 loss raises myristic acid and lowers VPS15 to worsen injury.","evidence":"NMT1 knockdown in rat ischemia-reperfusion model with MA injection, BCtDCS modulation, and infarct measurement","pmids":["40221008"],"confidence":"Medium","gaps":["Mechanism linking myristic acid accumulation to VPS15 downregulation unclear","Single in vivo model"]},{"year":2024,"claim":"Established host NMT1/NMT2 as required for mammarenavirus replication by myristoylating viral Z and SSP proteins, nominating NMT inhibition as a broad antiviral strategy.","evidence":"Pan-NMT inhibitor DDD85464, G2A mutation of Z/SSP, budding and fusion assays, and proteasome rescue (preprint)","pmids":[],"confidence":"Medium","gaps":["Preprint, not yet peer-reviewed","NMT1-specific versus NMT2-specific contributions not separated"]},{"year":null,"claim":"How NMT1 substrate selection, dual G/K modification, and downstream trafficking/stability outcomes are coordinated in a single cellular context, and how NMT1 is distinguished functionally from NMT2, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying model linking substrate choice to opposite stability outcomes","NMT1 vs NMT2 substrate division of labor undefined","No human structure-guided substrate prediction validated in cells"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,3,4,9]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[3,9]},{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[3,4,9]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[4,6,10]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[5]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[10]}],"complexes":[],"partners":["SORBS2","CHP1","ICAM-1","FBXO4","TMEM106B","VILIP3","PD-L1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P30419","full_name":"Glycylpeptide N-tetradecanoyltransferase 1","aliases":["Myristoyl-CoA:protein N-myristoyltransferase 1","HsNMT1","NMT 1","Type I N-myristoyltransferase","Peptide N-myristoyltransferase 1","Protein-lysine myristoyltransferase NMT1"],"length_aa":496,"mass_kda":56.8,"function":"Adds a myristoyl group to the N-terminal glycine residue of certain cellular and viral proteins (PubMed:22865860, PubMed:25255805, PubMed:32686708, PubMed:34999170, PubMed:9353336, PubMed:9506952). Also able to mediate N-terminal lysine myristoylation of proteins: catalyzes myristoylation of ARF6 on both 'Gly-2' and 'Lys-3' (PubMed:32103017, PubMed:32111831). Lysine myristoylation is required to maintain ARF6 on membranes during the GTPase cycle (PubMed:32103017)","subcellular_location":"Cytoplasm; Cytoplasm, cytosol; Membrane","url":"https://www.uniprot.org/uniprotkb/P30419/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/NMT1","classification":"Common Essential","n_dependent_lines":843,"n_total_lines":1208,"dependency_fraction":0.6978476821192053},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"FKBP5","stoichiometry":0.2},{"gene":"PSPC1","stoichiometry":0.2},{"gene":"RPS16","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/NMT1","total_profiled":1310},"omim":[{"mim_id":"620895","title":"GLUTAMATE-RICH PROTEIN 5; ERICH5","url":"https://www.omim.org/entry/620895"},{"mim_id":"620055","title":"PWP1 HOMOLOG, ENDONUCLEIN; PWP1","url":"https://www.omim.org/entry/620055"},{"mim_id":"603801","title":"N-MYRISTOYLTRANSFERASE 2; NMT2","url":"https://www.omim.org/entry/603801"},{"mim_id":"160993","title":"N-MYRISTOYLTRANSFERASE 1; NMT1","url":"https://www.omim.org/entry/160993"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cytosol","reliability":"Supported"},{"location":"Plasma membrane","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/NMT1"},"hgnc":{"alias_symbol":["NMT"],"prev_symbol":[]},"alphafold":{"accession":"P30419","domains":[{"cath_id":"3.40.630.170","chopping":"105-283","consensus_level":"medium","plddt":94.0341,"start":105,"end":283},{"cath_id":"3.40.630.170","chopping":"292-489","consensus_level":"medium","plddt":96.5209,"start":292,"end":489}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P30419","model_url":"https://alphafold.ebi.ac.uk/files/AF-P30419-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P30419-F1-predicted_aligned_error_v6.png","plddt_mean":83.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NMT1","jax_strain_url":"https://www.jax.org/strain/search?query=NMT1"},"sequence":{"accession":"P30419","fasta_url":"https://rest.uniprot.org/uniprotkb/P30419.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P30419/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P30419"}},"corpus_meta":[{"pmid":"2358444","id":"PMC_2358444","title":"nmt1 of fission yeast. A highly transcribed gene completely repressed by thiamine.","date":"1990","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/2358444","citation_count":752,"is_preprint":false},{"pmid":"8422997","id":"PMC_8422997","title":"TATA box mutations in the Schizosaccharomyces pombe nmt1 promoter affect transcription efficiency but not the transcription start point or thiamine repressibility.","date":"1993","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/8422997","citation_count":619,"is_preprint":false},{"pmid":"1594577","id":"PMC_1594577","title":"Comparative analysis of the beta transducin family with identification of several new members including PWP1, a nonessential gene of Saccharomyces cerevisiae that is divergently transcribed from NMT1.","date":"1992","source":"Proteins","url":"https://pubmed.ncbi.nlm.nih.gov/1594577","citation_count":100,"is_preprint":false},{"pmid":"12824382","id":"PMC_12824382","title":"Prediction of lipid posttranslational modifications and localization signals from protein sequences: big-Pi, NMT and PTS1.","date":"2003","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/12824382","citation_count":73,"is_preprint":false},{"pmid":"36617552","id":"PMC_36617552","title":"Blockade of NMT1 enzymatic activity inhibits N-myristoylation of VILIP3 protein and suppresses liver cancer progression.","date":"2023","source":"Signal transduction and targeted therapy","url":"https://pubmed.ncbi.nlm.nih.gov/36617552","citation_count":63,"is_preprint":false},{"pmid":"1429724","id":"PMC_1429724","title":"Studies of the catalytic activities and substrate specificities of Saccharomyces cerevisiae myristoyl-coenzyme A: protein N-myristoyltransferase deletion mutants and human/yeast Nmt chimeras in Escherichia coli and S. cerevisiae.","date":"1992","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/1429724","citation_count":49,"is_preprint":false},{"pmid":"34200747","id":"PMC_34200747","title":"Plasticity in Neuroblastoma Cell Identity Defines a Noradrenergic-to-Mesenchymal Transition (NMT).","date":"2021","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/34200747","citation_count":47,"is_preprint":false},{"pmid":"30446635","id":"PMC_30446635","title":"NMT1 inhibition modulates breast cancer progression through stress-triggered JNK pathway.","date":"2018","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/30446635","citation_count":37,"is_preprint":false},{"pmid":"9575206","id":"PMC_9575206","title":"Genetic and biochemical studies establish that the fungicidal effect of a fully depeptidized inhibitor of Cryptococcus neoformans myristoyl-CoA:protein N-myristoyltransferase (Nmt) is Nmt-dependent.","date":"1998","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9575206","citation_count":37,"is_preprint":false},{"pmid":"24248932","id":"PMC_24248932","title":"Characterization of a novel N-methyltransferase (NMT) from Catharanthus roseus plants : Detection of NMT and other enzymes of the indole alkaloid biosynthetic pathway in different cell suspension culture systems.","date":"1987","source":"Plant cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/24248932","citation_count":31,"is_preprint":false},{"pmid":"9606189","id":"PMC_9606189","title":"Nascent transcription from the nmt1 and nmt2 genes of Schizosaccharomyces pombe overlaps neighbouring genes.","date":"1998","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/9606189","citation_count":28,"is_preprint":false},{"pmid":"8613682","id":"PMC_8613682","title":"A radioresistant variant cell line, NMT-1R, isolated from a radiosensitive rat yolk sac tumour cell line, NMT-1: differences of early radiation-induced morphological changes, especially apoptosis.","date":"1996","source":"International journal of radiation biology","url":"https://pubmed.ncbi.nlm.nih.gov/8613682","citation_count":27,"is_preprint":false},{"pmid":"7992507","id":"PMC_7992507","title":"nmt2 of fission yeast: a second thiamine-repressible gene co-ordinately regulated with nmt1.","date":"1994","source":"Yeast (Chichester, England)","url":"https://pubmed.ncbi.nlm.nih.gov/7992507","citation_count":23,"is_preprint":false},{"pmid":"8163491","id":"PMC_8163491","title":"ntf1+ encodes a 6-cysteine zinc finger-containing transcription factor that regulates the nmt1 promoter in fission yeast.","date":"1994","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/8163491","citation_count":19,"is_preprint":false},{"pmid":"29777000","id":"PMC_29777000","title":"NMT1 and NMT3 N-Methyltransferase Activity Is Critical to Lipid Homeostasis, Morphogenesis, and Reproduction.","date":"2018","source":"Plant physiology","url":"https://pubmed.ncbi.nlm.nih.gov/29777000","citation_count":18,"is_preprint":false},{"pmid":"7507475","id":"PMC_7507475","title":"Establishment and characterization of a rat yolk sac tumor cell line, NMT-1, producing alpha-fetoprotein, with potential for lymphatic metastasis.","date":"1993","source":"Japanese journal of cancer research : Gann","url":"https://pubmed.ncbi.nlm.nih.gov/7507475","citation_count":15,"is_preprint":false},{"pmid":"34136404","id":"PMC_34136404","title":"N-Myristoylation by NMT1 Is POTEE-Dependent to Stimulate Liver Tumorigenesis via Differentially Regulating Ubiquitination of Targets.","date":"2021","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34136404","citation_count":12,"is_preprint":false},{"pmid":"21295003","id":"PMC_21295003","title":"A chemical compound for controlled expression of nmt1-driven gene in the fission yeast Schizosaccharomyces pombe.","date":"2011","source":"Analytical biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21295003","citation_count":10,"is_preprint":false},{"pmid":"36181773","id":"PMC_36181773","title":"Structural and Large-scale Analysis Unveil the Intertwined Paths Promoting NMT-catalyzed Lysine and Glycine Myristoylation.","date":"2022","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/36181773","citation_count":10,"is_preprint":false},{"pmid":"13680155","id":"PMC_13680155","title":"Characterization and expression of the Neurospora crassa nmt-1 gene.","date":"2003","source":"Current genetics","url":"https://pubmed.ncbi.nlm.nih.gov/13680155","citation_count":10,"is_preprint":false},{"pmid":"33805223","id":"PMC_33805223","title":"QSAR and Pharmacophore Modeling of Nitrogen Heterocycles as Potent Human N-Myristoyltransferase (Hs-NMT) Inhibitors.","date":"2021","source":"Molecules (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/33805223","citation_count":10,"is_preprint":false},{"pmid":"10470373","id":"PMC_10470373","title":"Expression of enzymes of covalent protein modification during regulated and dysregulated proliferation of mammary epithelial cells: PKA, PKC and NMT.","date":"1999","source":"Advances in enzyme regulation","url":"https://pubmed.ncbi.nlm.nih.gov/10470373","citation_count":9,"is_preprint":false},{"pmid":"37269961","id":"PMC_37269961","title":"NMT1 sustains ICAM-1 to modulate adhesion and migration of tumor cells.","date":"2023","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/37269961","citation_count":7,"is_preprint":false},{"pmid":"25801050","id":"PMC_25801050","title":"An IPTG-inducible derivative of the fission yeast nmt promoter.","date":"2015","source":"Yeast (Chichester, England)","url":"https://pubmed.ncbi.nlm.nih.gov/25801050","citation_count":7,"is_preprint":false},{"pmid":"37486304","id":"PMC_37486304","title":"Mechanism of Ligand Discrimination by the NMT1 Riboswitch.","date":"2023","source":"Journal of chemical information and modeling","url":"https://pubmed.ncbi.nlm.nih.gov/37486304","citation_count":6,"is_preprint":false},{"pmid":"34333213","id":"PMC_34333213","title":"Long noncoding RNA ANCR promotes migration, invasion, EMT progress and stemness of nasopharyngeal carcinoma cells via the miR-4731-5p/NMT1 axis.","date":"2021","source":"Pathology, research and practice","url":"https://pubmed.ncbi.nlm.nih.gov/34333213","citation_count":6,"is_preprint":false},{"pmid":"8533474","id":"PMC_8533474","title":"An 8.2 kb DNA segment from chromosome XIV carries the RPD3 and PAS8 genes as well as the Saccharomyces cerevisiae homologue of the thiamine-repressed nmt1 gene and a chromosome III-duplicated gene for a putative aryl-alcohol dehydrogenase.","date":"1995","source":"Yeast (Chichester, England)","url":"https://pubmed.ncbi.nlm.nih.gov/8533474","citation_count":6,"is_preprint":false},{"pmid":"31664711","id":"PMC_31664711","title":"Regulation of the thiamine pyrophosphate (TPP)-sensing riboswitch in NMT1 mRNA from Neurospora crassa.","date":"2019","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/31664711","citation_count":6,"is_preprint":false},{"pmid":"16874521","id":"PMC_16874521","title":"Joint regulation of the nmt1 promoter and sporulation by Thi1 and Thi5 in Schizosaccharomyces pombe.","date":"2006","source":"Current genetics","url":"https://pubmed.ncbi.nlm.nih.gov/16874521","citation_count":5,"is_preprint":false},{"pmid":"36002266","id":"PMC_36002266","title":"Identification of potential inhibitors for N-myristoyltransferase (NMT) protein of Plasmodium vivax.","date":"2022","source":"Journal of biomolecular structure & dynamics","url":"https://pubmed.ncbi.nlm.nih.gov/36002266","citation_count":5,"is_preprint":false},{"pmid":"35732157","id":"PMC_35732157","title":"NMT1 Enhances the Stemness of NSCLC Cells by Activating the PI3K/AKT Pathway.","date":"2022","source":"Pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/35732157","citation_count":3,"is_preprint":false},{"pmid":"40605065","id":"PMC_40605065","title":"Elucidating the role of N-myristoylation in the excessive membrane localization of PD-L1 in hypoxic cancers and developing a novel NMT1 inhibitor for combination with immune checkpoint blockade therapy.","date":"2025","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/40605065","citation_count":3,"is_preprint":false},{"pmid":"33937538","id":"PMC_33937538","title":"The role of NMT induction on odontogenic proliferation and differentiation of dental pulp stem cells.","date":"2021","source":"Heliyon","url":"https://pubmed.ncbi.nlm.nih.gov/33937538","citation_count":3,"is_preprint":false},{"pmid":"40451428","id":"PMC_40451428","title":"Myristoylation of TMEM106B by NMT1/2 regulates TMEM106B trafficking and turnover.","date":"2025","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/40451428","citation_count":2,"is_preprint":false},{"pmid":"38148048","id":"PMC_38148048","title":"TNF-α-induced Inhibition of Protein Myristoylation Via Binding Between NMT1 and Sorbs2 in Osteoblasts.","date":"2024","source":"In vivo (Athens, Greece)","url":"https://pubmed.ncbi.nlm.nih.gov/38148048","citation_count":1,"is_preprint":false},{"pmid":"36542915","id":"PMC_36542915","title":"Inspection of in-house designed novel thiochromone amino-acid conjugate derivatives as Lm-NMT inhibitor - An in-silico analysis.","date":"2022","source":"Journal of molecular graphics & modelling","url":"https://pubmed.ncbi.nlm.nih.gov/36542915","citation_count":1,"is_preprint":false},{"pmid":"40933604","id":"PMC_40933604","title":"Molecular docking and simulation analysis of selected herbal compounds against GP63, FPPS, and NMT, three important Leishmania major proteins.","date":"2025","source":"Research in pharmaceutical sciences","url":"https://pubmed.ncbi.nlm.nih.gov/40933604","citation_count":1,"is_preprint":false},{"pmid":"38975119","id":"PMC_38975119","title":"Development of a genetically encoded NMT indicator for detection of mercury ions based on the green fluorescent protein mNeonGreen and metallothionein II from rat liver.","date":"2024","source":"Heliyon","url":"https://pubmed.ncbi.nlm.nih.gov/38975119","citation_count":1,"is_preprint":false},{"pmid":"31161515","id":"PMC_31161515","title":"N-Myristoyl Transferase (NMT)-Catalyzed Labeling of Bacterial Proteins for Imaging in Fixed and Live Cells.","date":"2019","source":"Methods in molecular biology (Clifton, N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/31161515","citation_count":1,"is_preprint":false},{"pmid":"40221008","id":"PMC_40221008","title":"Identification of NMT1/MA/VPS15 signal pathway as potential therapeutic target in rat cerebral ischemia injury.","date":"2025","source":"Experimental neurology","url":"https://pubmed.ncbi.nlm.nih.gov/40221008","citation_count":0,"is_preprint":false},{"pmid":"41406521","id":"PMC_41406521","title":"Mitigating cadmium toxicity in rice through tandem application of zinc oxide nanoparticles and Serendipita indica as revealed by multi-omics and NMT-based ion flux analysis.","date":"2025","source":"Journal of hazardous materials","url":"https://pubmed.ncbi.nlm.nih.gov/41406521","citation_count":0,"is_preprint":false},{"pmid":"24908050","id":"PMC_24908050","title":"The crystal structure of pyrimidine/thiamin biosynthesis precursor-like domain-containing protein CAE31940 from proteobacterium Bordetella bronchiseptica RB50, and evolutionary insight into the NMT1/THI5 family.","date":"2014","source":"Journal of structural and functional genomics","url":"https://pubmed.ncbi.nlm.nih.gov/24908050","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.08.01.606235","title":"Cellular N-myristoyl transferases Are Required for Mammarenavirus Multiplication","date":"2024-08-01","source":"bioRxiv","url":"https://doi.org/10.1101/2024.08.01.606235","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.06.20.660686","title":"N-myristoyltransferase Inhibitors as Candidate Broad-Spectrum Antivirals to Treat Viral Infections Promoted by Immunosuppression Associated with JAK Inhibitors Therapy","date":"2025-06-21","source":"bioRxiv","url":"https://doi.org/10.1101/2025.06.20.660686","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.06.22.600193","title":"An improved tetracycline-inducible expression system for fission yeast","date":"2024-06-27","source":"bioRxiv","url":"https://doi.org/10.1101/2024.06.22.600193","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.07.10.663842","title":"Fully Automated and Scalable Pipeline for Macaque Brain Registration","date":"2025-07-16","source":"bioRxiv","url":"https://doi.org/10.1101/2025.07.10.663842","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.11.22.624841","title":"diaPASEF-Powered Chemoproteomics Enables Deep Kinome Interaction Profiling","date":"2024-11-22","source":"bioRxiv","url":"https://doi.org/10.1101/2024.11.22.624841","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.03.24.644888","title":"MolAI: A Deep Learning Framework for Data-driven Molecular Descriptor Generation and Advanced Drug Discovery Applications","date":"2025-03-27","source":"bioRxiv","url":"https://doi.org/10.1101/2025.03.24.644888","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.01.07.631647","title":"Micromorphology and ammonium transmembrane transport characteristics in roots of rice seedlings","date":"2025-01-10","source":"bioRxiv","url":"https://doi.org/10.1101/2025.01.07.631647","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.07.28.605509","title":"Parallel evolution of methyltransferases leads to vobasine biosynthesis in  <i>Tabernaemontana elegans</i>  and  <i>Catharanthus roseus</i>","date":"2024-07-29","source":"bioRxiv","url":"https://doi.org/10.1101/2024.07.28.605509","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":25219,"output_tokens":3917,"usd":0.067206,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11670,"output_tokens":4299,"usd":0.082913,"stage2_stop_reason":"end_turn"},"total_usd":0.150119,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1992,\n      \"finding\": \"Saccharomyces cerevisiae Nmt1p (NMT1 ortholog) is an essential, monomeric 455-residue enzyme whose minimal catalytic domain lies between Ile59–Phe96 and Gly451–Leu455, as determined by deletion mutagenesis. The amino-terminal 59 residues play a non-catalytic targeting role, allowing the cytosolic enzyme to access myristoyl-CoA pools derived from exogenous fatty acid activation by acyl-CoA synthetases.\",\n      \"method\": \"Deletion mutagenesis of Nmt1p expressed in E. coli co-expression system; in vitro enzyme assay with [3H]myristate incorporation; rescue of nmt1 null allele in yeast\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro enzyme assays plus mutagenesis plus genetic complementation, multiple orthogonal methods in one rigorous study\",\n      \"pmids\": [\"1429724\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1992,\n      \"finding\": \"Structural elements determining substrate specificity differences between yeast Nmt1p and human NMT were mapped using chimeric enzymes: recognition of the human-specific substrate Gz-alpha involves elements distributed from the amino-terminal half through the region Leu352–Lys410 of the 416-residue human enzyme, and formation of fully functional peptide- and myristoyl-CoA-binding sites requires contributions from both amino- and carboxyl-terminal halves of either enzyme.\",\n      \"method\": \"E. coli co-expression of chimeric human/yeast NMT constructs with substrate G-protein alpha subunits; [3H]myristate incorporation assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — systematic chimeric enzyme mutagenesis with functional reconstitution assay, multiple chimeras tested\",\n      \"pmids\": [\"1429724\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"Cryptococcus neoformans NMT (Nmt) is essential for fungal viability; under-myristoylation of an ADP ribosylation factor (Arf) substrate contributes to a temperature-sensitive myristic acid auxotrophy phenotype caused by a conditional-lethal nmt allele. A depeptidized inhibitor exerted fungicidal effects in an Nmt-dependent manner, with Ki(app) of 1.8 µM for fungal Nmt and 9 µM for human NMT.\",\n      \"method\": \"Homologous recombination to introduce conditional-lethal NMT allele; isogenic strain comparison with human NMT rescue; kinetic inhibition studies with purified fungal and human Nmt; protein synthesis inhibition assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — genetic epistasis with biochemical rescue, purified-enzyme kinetics, and strain-specific pharmacology, multiple orthogonal methods\",\n      \"pmids\": [\"9575206\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NMT (N-myristoyltransferase) catalyzes both G-myristoylation (N-terminal glycine α-amino group) and K-myristoylation (lysine ε-amino group). Crystal structure and kinetic studies show that K-myristoylation uses direct interactions between the substrate's reactive amino group and the NMT catalytic base, whereas G-myristoylation uses a water-mediated interaction, making G-myristoylation more efficient. K-myristoylation motifs are depleted in humans, suggesting evolutionary pressure against it, and K-myristoylation arises only from post-translational events.\",\n      \"method\": \"Crystallography, mass spectrometry, in vitro kinetic assays, in silico analysis\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure combined with kinetics and MS in one study, multiple orthogonal methods\",\n      \"pmids\": [\"36181773\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"NMT1 myristoylates VILIP3 (Visinin-like protein 3) as a substrate. Desloratadine binds to Asn-246 in NMT1 and inhibits its enzymatic activity, blocking NMT1-mediated myristoylation of VILIP3 and disrupting downstream NFκB/Bcl-2 signaling in hepatocellular carcinoma cells.\",\n      \"method\": \"Drug affinity responsive target stability (DARTS) and surface plasmon resonance (SPR) to identify NMT1 as desloratadine target; metabolic labeling and mass spectrometry to identify VILIP3 as NMT1 substrate; NMT1 knockdown and overexpression in vitro and in vivo (CDX, PDO, PDX models)\",\n      \"journal\": \"Signal transduction and targeted therapy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal target identification by SPR and DARTS, metabolic labeling MS substrate identification, in vivo models, multiple orthogonal methods\",\n      \"pmids\": [\"36617552\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Genetic inhibition of NMT1 in breast cancer cells promotes oxidative stress (ROS) and ER stress, which cross-talk with mitochondria to produce more ROS; both stresses activate the JNK pathway, leading to autophagy that suppresses breast cancer progression. ROS negatively regulates NMT1 expression and NMT1 knockdown conversely promotes oxidative stress, forming a feedback loop.\",\n      \"method\": \"NMT1 knockdown (siRNA/shRNA) in breast cancer cell lines; in vitro proliferation/invasion assays; in vivo xenograft models; ROS measurements; JNK pathway western blotting; autophagy assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KD with defined cellular phenotype and pathway placement, single lab, multiple assays\",\n      \"pmids\": [\"30446635\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NMT1-mediated N-myristoylation differentially regulates two categories of protein targets in liver cancer: NDP proteins (LXN, RPL29, FAU) are N-myristoylated by NMT1 in a POTEE-dependent manner causing their destabilization via RPL7A/HIST1H4H ubiquitin E3 ligase-mediated ubiquitination, whereas NUP proteins (AHSG, ALB, TF) are N-myristoylated by NMT1, increasing their stability by preventing ubiquitination through HBB. Both RPL7A and HBB functions are NMT1-dependent.\",\n      \"method\": \"Liver-conditional NMT1 knockout mice; iTRAQ proteomics; click chemistry assay for N-myristoylation levels; parallel reaction monitoring (PRM); co-immunoprecipitation; ubiquitination assays\",\n      \"journal\": \"Frontiers in oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional KO mouse model plus proteomics plus click chemistry, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"34136404\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"NMT1 sustains ICAM-1 protein levels by preventing its ubiquitination and proteasomal degradation; mechanistically, NMT1 myristoylates the N-terminus of ICAM-1 and inhibits the Ub E3 ligase F-box protein 4 (FBXO4), prolonging the half-life of ICAM-1 protein and thereby sustaining cell adhesion while suppressing tumor cell migration.\",\n      \"method\": \"NMT1 knockdown; co-immunoprecipitation; ubiquitination assays; half-life assays; migration/adhesion functional assays\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KD with defined phenotype plus mechanistic co-IP and ubiquitination assays, single lab\",\n      \"pmids\": [\"37269961\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"TNF-α stimulation in osteoblasts (MC3T3-E1 cells) increases NMT1 expression but inhibits overall protein myristoylation; immunoprecipitation and mass spectrometry identified Sorbs2 as a novel NMT1-binding protein, and the NMT1–Sorbs2 interaction upon TNF-α stimulation is responsible for inhibition of myristoylation.\",\n      \"method\": \"Click-it assay for myristoylated proteins; RNA interference; mass spectrometry; immunoprecipitation; immunocytochemistry; western blotting\",\n      \"journal\": \"In vivo (Athens, Greece)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — reciprocal Co-IP plus MS identification of binding partner plus click chemistry assay, single lab\",\n      \"pmids\": [\"38148048\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NMT1 myristoylates TMEM106B at its glycine-2 α-amino group and lysine-3 ε-amino group. Myristoylation promotes lysosomal degradation of TMEM106B, regulates generation of its C-terminal fragments, and controls its trafficking: non-myristoylated TMEM106B accumulates on the cell surface instead of being trafficked to lysosomes.\",\n      \"method\": \"NMT1/2 enzymatic assays; site-directed mutagenesis (G2A, K3 mutants); subcellular fractionation; live cell imaging; lysosomal inhibitor experiments; western blotting for TMEM106B levels and CTFs\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — enzymatic myristoylation assay with mutagenesis, trafficking studies, and degradation assays, multiple orthogonal methods in one study\",\n      \"pmids\": [\"40451428\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"HIF1α upregulates NMT1, which mediates myristoylation of calcineurin B homologous protein 1 (CHP1) under hypoxia. Myristoylated CHP1 binds PD-L1 and facilitates its rapid translocation to the cell membrane, enhancing PD-L1-mediated immune evasion. The NMT1 inhibitor PCLX-001 blocks CHP1 myristoylation, disrupting excessive PD-L1 membrane localization.\",\n      \"method\": \"NMT1 inhibitor (PCLX-001) treatment; HIF1α overexpression/knockdown; co-immunoprecipitation of CHP1 and PD-L1; flow cytometry for PD-L1 membrane localization; xenograft mouse models\",\n      \"journal\": \"Journal of experimental & clinical cancer research : CR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP of binding partners, pharmacological inhibition, localization assays, in vivo model, single lab\",\n      \"pmids\": [\"40605065\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NMT1 downregulation in cerebral ischemia leads to accumulation of myristic acid (MA) in the penumbra, which in turn reduces VPS15 (phosphoinositide 3-kinase regulatory subunit 4) expression, exacerbating ischemia injury. NMT1 thus acts on MA to regulate VPS15 expression, defining an NMT1/MA/VPS15 signaling pathway.\",\n      \"method\": \"NMT1 knockdown in rat cerebral ischemia-reperfusion model; intraperitoneal MA injection; BCtDCS to modulate MA levels; western blotting for NMT1 and VPS15; infarct volume measurement\",\n      \"journal\": \"Experimental neurology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo knockdown with defined pathway placement and rescue experiments, single lab\",\n      \"pmids\": [\"40221008\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Host cell NMT1 (and NMT2) are required for mammarenavirus multiplication: the validated pan-NMT inhibitor DDD85464 exerts antiviral activity against LCMV, JUNV, and LASV, correlating with reduced Z protein budding activity, reduced GP2-mediated fusion activity, and proteasome-mediated degradation of the Z protein. The Z protein and stable signal peptide (SSP) of mammarenaviruses are N-terminally myristoylated by host NMT1/NMT2.\",\n      \"method\": \"Pan-NMT inhibitor DDD85464 treatment; G2A mutation analysis of viral Z and SSP proteins; virus budding assays; GP2-mediated fusion assays; proteasome inhibitor rescue experiments\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological inhibition with on-target validation, multiple viral substrates and functional assays, preprint not yet peer-reviewed\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"Human NMT1 is an essential monomeric N-myristoyltransferase that co- and post-translationally transfers myristate from myristoyl-CoA to N-terminal glycine residues (and, less efficiently, to lysine ε-amino groups) of substrate proteins including VILIP3, CHP1, ICAM-1, TMEM106B, and viral Z/SSP proteins; its catalytic activity—requiring contributions from both N- and C-terminal halves for substrate and acyl-CoA binding—controls substrate stability (via modulation of ubiquitin-mediated degradation), subcellular trafficking (e.g., directing TMEM106B to lysosomes and CHP1/PD-L1 to the plasma membrane), and downstream signaling (NFκB/Bcl-2, JNK/autophagy, PI3K/AKT, NMT1/MA/VPS15), with its activity inhibitable at Asn-246 by small molecules such as desloratadine and PCLX-001.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"NMT1 is an essential N-myristoyltransferase that transfers myristate from myristoyl-CoA onto the N-terminal glycine α-amino group of substrate proteins, a co- and post-translational lipid modification that governs substrate stability, trafficking, and downstream signaling [#0, #4]. The enzyme is monomeric, with a minimal catalytic domain whose substrate- and acyl-CoA-binding sites require functional contributions from both its amino- and carboxyl-terminal halves [#0, #1]; structural and kinetic work shows it preferentially catalyzes glycine myristoylation through a water-mediated chemistry while also performing less efficient lysine ε-amino myristoylation via direct interaction with the catalytic base [#3]. Through these modifications NMT1 controls the fate of diverse substrates: it directs membrane and trafficking outcomes, sending myristoylated TMEM106B to lysosomes for degradation while non-myristoylated TMEM106B accumulates at the cell surface [#9], and promoting CHP1-dependent translocation of PD-L1 to the plasma membrane to enable immune evasion under hypoxia [#10]. NMT1 also acts as a stability switch by modulating ubiquitin-mediated degradation—stabilizing ICAM-1 by inhibiting the E3 ligase FBXO4 [#7] and differentially destabilizing or stabilizing distinct protein classes in liver cancer through E3-ligase-dependent ubiquitination [#6]. Beyond protein targets, NMT1 shapes signaling through its product and substrate networks, regulating ROS/JNK-driven autophagy [#5], NF\\u03baB/Bcl-2 signaling via VILIP3 [#4], and an NMT1/myristic-acid/VPS15 axis in cerebral ischemia [#11]. Its catalytic activity is druggable at Asn-246 by small molecules including desloratadine and by pan-NMT inhibitors such as PCLX-001 and DDD85464, the latter blocking myristoylation of viral Z and stable-signal-peptide proteins required for mammarenavirus multiplication [#4, #10, #12].\",\n  \"teleology\": [\n    {\n      \"year\": 1992,\n      \"claim\": \"Established that the N-myristoyltransferase is an essential monomeric enzyme with a defined catalytic core and an N-terminal targeting region, answering what minimal structure supports myristoyl transfer.\",\n      \"evidence\": \"Deletion mutagenesis of yeast Nmt1p in E. coli co-expression with in vitro [3H]myristate assays and rescue of an nmt1 null allele\",\n      \"pmids\": [\"1429724\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Defined in yeast ortholog rather than human NMT1\", \"Does not resolve atomic-level catalytic mechanism\"]\n    },\n    {\n      \"year\": 1992,\n      \"claim\": \"Mapped how substrate specificity is encoded, showing functional peptide- and acyl-CoA-binding sites require both enzyme halves and explaining species-specific substrate recognition.\",\n      \"evidence\": \"Chimeric human/yeast NMT constructs co-expressed with G-protein alpha substrates and assayed by [3H]myristate incorporation\",\n      \"pmids\": [\"1429724\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specificity elements mapped by chimeras, not full structure\", \"Limited to a small set of test substrates\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"Demonstrated NMT essentiality for viability in a pathogen and validated it as a druggable antifungal target, establishing the enzyme as therapeutically tractable.\",\n      \"evidence\": \"Conditional-lethal NMT allele in Cryptococcus neoformans with human NMT rescue and purified-enzyme inhibitor kinetics\",\n      \"pmids\": [\"9575206\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Demonstrated in fungal system\", \"Human NMT inhibition shown only kinetically, not in cells\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Resolved the chemical basis for dual glycine versus lysine myristoylation, explaining why N-terminal glycine modification dominates while lysine modification is rare and post-translational.\",\n      \"evidence\": \"Crystallography combined with in vitro kinetics, mass spectrometry, and in silico motif analysis\",\n      \"pmids\": [\"36181773\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Biological scope of K-myristoylation in cells not defined\", \"Specific human substrates not enumerated here\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Placed NMT1 within an oxidative/ER-stress and JNK-autophagy axis, defining a tumor-suppressive consequence of NMT1 loss in breast cancer.\",\n      \"evidence\": \"NMT1 knockdown in breast cancer lines with ROS measurement, JNK western blotting, autophagy assays, and xenografts\",\n      \"pmids\": [\"30446635\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct myristoylation substrate driving the phenotype not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Showed NMT1 myristoylation bidirectionally controls substrate stability via E3-ligase-mediated ubiquitination, distinguishing destabilized from stabilized target classes in liver cancer.\",\n      \"evidence\": \"Liver-conditional NMT1 knockout mice with iTRAQ proteomics, click chemistry, PRM, co-IP, and ubiquitination assays\",\n      \"pmids\": [\"34136404\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which myristoylation directs opposite ubiquitination outcomes unclear\", \"POTEE/HBB roles not structurally defined\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified VILIP3 as an NMT1 substrate and pinned the drug-binding site at Asn-246, connecting catalytic inhibition to NF\\u03baB/Bcl-2 signaling and providing a defined pharmacophore.\",\n      \"evidence\": \"DARTS and SPR target identification, metabolic labeling MS for substrate, and NMT1 perturbation across CDX/PDO/PDX models\",\n      \"pmids\": [\"36617552\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct link from VILIP3 myristoylation to NF\\u03baB mechanism not fully resolved\", \"Selectivity of desloratadine over NMT2 not addressed\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined NMT1 as a stabilizer of ICAM-1 by inhibiting the E3 ligase FBXO4, linking myristoylation to cell adhesion and migration control.\",\n      \"evidence\": \"NMT1 knockdown with co-IP, ubiquitination, half-life, and migration/adhesion assays\",\n      \"pmids\": [\"37269961\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether NMT1 inhibits FBXO4 directly or via myristoylation unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified Sorbs2 as an NMT1-binding partner that suppresses global myristoylation under TNF-\\u03b1, revealing inflammatory regulation of NMT1 activity in osteoblasts.\",\n      \"evidence\": \"Click-it myristoylation assay, RNAi, immunoprecipitation, and mass spectrometry in MC3T3-E1 cells\",\n      \"pmids\": [\"38148048\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism by which Sorbs2 inhibits NMT1 not defined\", \"Single lab and single cell model\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Showed NMT1 myristoylates TMEM106B at both glycine-2 and lysine-3, controlling its lysosomal degradation, fragment generation, and surface-versus-lysosome trafficking.\",\n      \"evidence\": \"NMT1/2 enzymatic assays with G2A/K3 mutants, subcellular fractionation, live imaging, and lysosomal inhibitor experiments\",\n      \"pmids\": [\"40451428\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative in vivo contributions of NMT1 versus NMT2 not separated\", \"Disease relevance of TMEM106B fragments not tested here\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Connected HIF1\\u03b1-driven NMT1 to CHP1 myristoylation and PD-L1 membrane trafficking, defining a hypoxia-induced immune-evasion mechanism druggable by PCLX-001.\",\n      \"evidence\": \"PCLX-001 treatment, HIF1\\u03b1 perturbation, CHP1/PD-L1 co-IP, flow cytometry for PD-L1, and xenografts\",\n      \"pmids\": [\"40605065\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct structural basis of myristoylated-CHP1\\u2013PD-L1 interaction unresolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined an NMT1/myristic-acid/VPS15 axis in cerebral ischemia, showing NMT1 loss raises myristic acid and lowers VPS15 to worsen injury.\",\n      \"evidence\": \"NMT1 knockdown in rat ischemia-reperfusion model with MA injection, BCtDCS modulation, and infarct measurement\",\n      \"pmids\": [\"40221008\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking myristic acid accumulation to VPS15 downregulation unclear\", \"Single in vivo model\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Established host NMT1/NMT2 as required for mammarenavirus replication by myristoylating viral Z and SSP proteins, nominating NMT inhibition as a broad antiviral strategy.\",\n      \"evidence\": \"Pan-NMT inhibitor DDD85464, G2A mutation of Z/SSP, budding and fusion assays, and proteasome rescue (preprint)\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, not yet peer-reviewed\", \"NMT1-specific versus NMT2-specific contributions not separated\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How NMT1 substrate selection, dual G/K modification, and downstream trafficking/stability outcomes are coordinated in a single cellular context, and how NMT1 is distinguished functionally from NMT2, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unifying model linking substrate choice to opposite stability outcomes\", \"NMT1 vs NMT2 substrate division of labor undefined\", \"No human structure-guided substrate prediction validated in cells\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 3, 4, 9]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [3, 9]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [3, 4, 9]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [4, 6, 10]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [10]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"Sorbs2\",\n      \"CHP1\",\n      \"ICAM-1\",\n      \"FBXO4\",\n      \"TMEM106B\",\n      \"VILIP3\",\n      \"PD-L1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}