{"gene":"STK10","run_date":"2026-06-10T07:46:43","timeline":{"discoveries":[{"year":1997,"finding":"LOK (STK10) is a serine/threonine kinase (not tyrosine kinase) that autophosphorylates and phosphorylates myelin basic protein and histone IIA; it belongs to the STE20 family with an N-terminal kinase domain and C-terminal coiled-coil/proline-rich region; it does not activate ERK, JNK, or p38 MAP kinases when co-expressed in COS7 cells.","method":"In vitro kinase assay, co-expression in COS7 cells, Western blot","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — in vitro kinase assay with substrate phosphorylation, single lab, foundational characterization paper","pmids":["9278426"],"is_preprint":false},{"year":2000,"finding":"LOK-deficient mice show enhanced LFA-1 clustering and accelerated LFA-1/ICAM-mediated T cell aggregation upon mitogen stimulation, without changes in total LFA-1 or ICAM levels, indicating LOK negatively regulates LFA-1 clustering and lymphocyte adhesion.","method":"LOK knockout mouse model, soluble ICAM-1 binding assay, flow cytometry","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — genetic KO with defined cellular phenotype, single lab, single method","pmids":["10692593"],"is_preprint":false},{"year":2002,"finding":"Overexpression of LOK attenuates MEKK1-induced and Raji/SEE-induced CD28RE/AP1 reporter gene activation and IL-2 production in Jurkat T cells, indicating LOK opposes MEKK1 in the CD28 signaling pathway.","method":"Luciferase reporter assay, co-transfection in Jurkat cells, IL-2 measurement","journal":"The Biochemical journal","confidence":"Low","confidence_rationale":"Tier 3 / Weak — overexpression reporter assay, single lab, no direct biochemical mechanism established","pmids":["11903060"],"is_preprint":false},{"year":2003,"finding":"STK10 (human LOK) associates with PLK1 in cells and phosphorylates PLK1 in vitro; dominant-negative STK10 expression in NIH-3T3 cells causes increased DNA content, suggesting STK10 functions as a polo-like kinase kinase regulating PLK1.","method":"Co-immunoprecipitation, in vitro kinase assay, engineered NIH-3T3 cell lines with flow cytometry cell cycle analysis","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP plus in vitro kinase assay plus dominant-negative cell phenotype, single lab","pmids":["12639966"],"is_preprint":false},{"year":2009,"finding":"LOK is a major ERM kinase in lymphocytes: it is enriched at the plasma membrane near ERM proteins, directly phosphorylates moesin at its C-terminal threonine in vitro with preferential specificity (including unusual preference for Tyr at P-2), and LOK knockout mice show >50% reduction in ERM phosphorylation; loss of LOK enhances lymphocyte migration and polarization in response to chemokine.","method":"Mass spectrometry localization, immunofluorescence, in vitro peptide kinase assay, LOK kinase domain transfection, LOK knockout mouse model","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro kinase assay with substrate specificity profiling, genetic KO with defined phosphorylation and functional phenotype, multiple orthogonal methods","pmids":["19255442"],"is_preprint":false},{"year":2012,"finding":"LOK and SLK are the relevant kinases driving apical restriction of ezrin in polarized epithelial cells; both kinases are enriched in microvilli and locally activated there; drug-resistant LOK/SLK variants are sufficient to restrict ezrin to the apical domain, while expression of their regulatory regions inhibits local ezrin phosphorylation by endogenous kinases.","method":"Proteomic approaches, RNAi knockdown, drug-resistant kinase variants, immunofluorescence localization","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — RNAi plus drug-resistant rescue variants plus localization studies, multiple orthogonal methods in one study","pmids":["23209304"],"is_preprint":false},{"year":2013,"finding":"Wild-type STK10 suppresses NF-κB activity and potentiates dexamethasone-induced apoptosis; PTCL-associated missense mutations (R634H, L85P, K277E) reduce this pro-apoptotic activity, with L85P and K277E having more profound anti-apoptotic effects than R634H.","method":"NF-κB reporter assay, apoptosis assay (dexamethasone), site-directed mutagenesis, transfection","journal":"Oncology reports","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — functional assays with mutagenesis, single lab, no direct biochemical target of NF-κB regulation identified","pmids":["23842845"],"is_preprint":false},{"year":2016,"finding":"In Drosophila GSCs, DNA damage activates Lok kinase, which is required for GSC loss and progeny differentiation defects; elimination of Lok or its kinase activity rescues these phenotypes; Lok-dependent signaling decreases expression of differentiation factor Bam.","method":"Genetic epistasis (lok knockout/kinase-dead), heat-shock I-CreI endonuclease and X-ray irradiation, immunofluorescence, Drosophila ovary model","journal":"Development (Cambridge, England)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with multiple DNA-damage inducers and kinase-dead allele, Drosophila ortholog","pmids":["27729408"],"is_preprint":false},{"year":2017,"finding":"LOK activates ezrin through a multi-step mechanism: (1) PIP2 binding to ezrin induces a conformational change; (2) the LOK C-terminal domain inserts to wedge apart the ezrin membrane- and F-actin-binding domains; (3) the LOK N-terminal kinase domain accesses a site 40 residues distal from the consensus sequence to phosphorylate the correct threonine. This ensures ezrin is only phosphorylated at the plasma membrane.","method":"In vitro reconstitution system, biochemical domain-mapping, mutagenesis, lipid-binding assays","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with mechanistic dissection, mutagenesis, and domain-mapping, multiple orthogonal biochemical methods in one study","pmids":["28430576"],"is_preprint":false},{"year":2021,"finding":"STK10 knockout in prostate cancer DU145 cells inhibits cell migration and promotes proliferation; these effects are mediated via inhibition of p38 MAPK activation and reduced ERM protein phosphorylation.","method":"CRISPR-Cas9 knockout, Western blot (phospho-ERM, phospho-p38), migration and proliferation assays","journal":"Experimental and therapeutic medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — CRISPR KO with defined phospho-signaling readouts and cellular phenotypes, single lab","pmids":["34149897"],"is_preprint":false},{"year":2021,"finding":"Crystal structures of SLK and STK10 with maleimide-scaffold inhibitors were determined, revealing the binding mode and structural basis for selectivity between SLK and STK10; cellular target engagement assays confirmed inhibitor binding to STK10 in cells.","method":"X-ray crystallography, cellular target engagement assay (NanoBRET or similar), medicinal chemistry SAR","journal":"Journal of medicinal chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure with cellular validation, multiple inhibitor series with defined binding modes","pmids":["34463505"],"is_preprint":false},{"year":2022,"finding":"Host Stk10 knockout in mice results in increased tumor growth associated with decreased activated/effector cytotoxic T lymphocytes and increased vessel density in the tumor microenvironment, indicating STK10 modulates anti-tumor immunity through CTL activity and angiogenesis regulation.","method":"Stk10 knockout mouse model, tumor implantation assay, immunofluorescence/flow cytometry of tumor-infiltrating immune cells","journal":"Biology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — genetic KO with in vivo tumor model and cellular immune phenotyping, single lab","pmids":["36421382"],"is_preprint":false},{"year":2024,"finding":"Knockdown of STK10 in K562 cells inhibits erythroid differentiation and promotes apoptosis, associated with inhibition of ribosome biogenesis, reduced ribosome levels, and activation of the p53 signaling pathway.","method":"shRNA knockdown in K562 cells, erythroid differentiation assay, ribosome profiling, Western blot (p53 pathway)","journal":"Annals of hematology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — knockdown with phenotype and pathway readouts, single lab, no direct biochemical mechanism linking STK10 to ribosome biogenesis","pmids":["38761185"],"is_preprint":false},{"year":2025,"finding":"A co-crystal structure of a macrocyclic inhibitor bound to STK10 revealed a unique back-pocket binding mode distinct from SLK, providing structural basis for selective STK10 inhibition; compound 23 showed nanomolar STK10 activity in cells.","method":"X-ray co-crystallography, biophysical assays, cellular activity assays","journal":"ACS medicinal chemistry letters","confidence":"High","confidence_rationale":"Tier 1 / Weak — crystal structure with cellular validation, single study but direct structural evidence","pmids":["41256983"],"is_preprint":false},{"year":2026,"finding":"STK10 in platelets is phosphorylated upon activation; platelet-specific STK10 knockout mice show impaired hemostasis, reduced platelet aggregation, α-granule release, αIIbβ3 activation, procoagulant activity, spreading, and clot retraction; STK10 directly phosphorylates integrin-linked kinase (ILK) at Ser343 as identified by immunoprecipitation-mass spectrometry and confirmed by in vitro phosphorylation assay; upstream, STK10 phosphorylation is regulated by calcium, PKC, and PI3K signaling.","method":"Platelet-specific conditional KO mice, quantitative phosphoproteomics, immunoprecipitation-mass spectrometry, in vitro kinase assay, platelet functional assays","journal":"Blood","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro kinase assay confirming direct ILK phosphorylation, conditional KO with multiple functional readouts, phosphoproteomics, multiple orthogonal methods","pmids":["41055696"],"is_preprint":false},{"year":2026,"finding":"The LOK C-terminal domain (LOK-CTD) mediates colocalization of LOK with ezrin at the apical surface of epithelial cells; the LOK-CTD forms dimers, binds negatively charged phospholipids, and shares structural similarity to inverse BAR (IBAR) domains; these properties are required for LOK-ezrin colocalization.","method":"Biochemical assays (lipid binding, dimerization), predictive bioinformatics, molecular dynamics simulations (atomistic and coarse-grained), human cell-based colocalization assays","journal":"Biophysical journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — biochemical assays plus cell-based functional assays plus MD simulations, single lab, multiple orthogonal methods","pmids":["41958020"],"is_preprint":false},{"year":2026,"finding":"Platelet STK10 deletion reduces deep vein thrombus formation in mice; STK10 phosphorylation and ILK (Ser343) phosphorylation are increased in platelets during DVT development; STK10 KO inhibits platelet-neutrophil interactions, NET formation, and platelet procoagulant activity in venous thrombi.","method":"Platelet-specific STK10 KO mice, inferior vena cava ligation DVT model, immunofluorescence, in vitro NET assay, phosphoprotein analysis","journal":"Journal of thrombosis and haemostasis","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — conditional KO in vivo model with multiple cellular readouts, single lab, extends findings from companion arterial thrombosis paper","pmids":["41791658"],"is_preprint":false},{"year":2025,"finding":"ERK phosphorylates the C-terminal tail of LOK, inhibiting LOK's activation of Ezrin in the cell body; this releases Ezrin's inhibition of Rho (via ARHGAP18) and promotes stress fiber assembly and cell migration, placing LOK in an ERK→LOK→Ezrin→ARHGAP18→Rho pathway.","method":"Cell-based phosphorylation assays, genetic epistasis, Rho activity assays, stress fiber imaging, Ezrin activity assays","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — genetic epistasis and cell-based biochemical assays, preprint, single lab, not yet peer-reviewed","pmids":["bio_10.1101_2025.11.15.688645"],"is_preprint":true}],"current_model":"STK10/LOK is a STE20-family serine/threonine kinase that primarily functions as a major ERM (ezrin/radixin/moesin) kinase enriched at the plasma membrane, where it phosphorylates ERM C-terminal threonines through a PIP2-dependent, multi-step wedge mechanism involving its C-terminal IBAR-like domain; it regulates lymphocyte migration, polarization, and LFA-1-mediated adhesion; acts as a PLK1-activating kinase; directly phosphorylates ILK at Ser343 in platelets to regulate arterial and venous thrombosis; suppresses NF-κB and promotes apoptosis; and is inhibited by ERK-mediated phosphorylation of its C-terminal tail to release Ezrin's inhibition of Rho for stress fiber-driven cell migration."},"narrative":{"mechanistic_narrative":"STK10 (LOK) is a STE20-family serine/threonine kinase whose principal characterized function is acting as a major ERM (ezrin/radixin/moesin) kinase that couples plasma-membrane signaling to cortical actin organization, cell adhesion, and migration [PMID:19255442, PMID:23209304]. It autophosphorylates and uses an N-terminal kinase domain to directly phosphorylate the regulatory C-terminal threonine of ERM proteins with distinctive substrate specificity, and its loss reduces ERM phosphorylation by more than half in lymphocytes while enhancing chemokine-driven migration and polarization [PMID:9278426, PMID:19255442]. STK10 achieves spatial fidelity through a multi-step wedge mechanism in which PIP2 binding primes ezrin, the kinase's C-terminal domain inserts to separate the membrane- and actin-binding lobes, and the kinase domain then accesses a non-consensus phosphosite, ensuring phosphorylation occurs only at the membrane [PMID:28430576]; the same C-terminal domain dimerizes, binds anionic phospholipids, resembles an inverse-BAR domain, and drives apical colocalization with ezrin in epithelial cells [PMID:41958020, PMID:23209304]. Beyond ERM regulation, STK10 negatively regulates LFA-1 clustering and lymphocyte adhesion [PMID:10692593], associates with and phosphorylates PLK1 as a polo-like kinase kinase [PMID:12639966], and in platelets is activated downstream of calcium/PKC/PI3K signaling to directly phosphorylate integrin-linked kinase (ILK) at Ser343, controlling aggregation, granule release, integrin activation, and both arterial-type and deep-vein thrombosis [PMID:41055696, PMID:41791658]. STK10 also suppresses NF-κB activity and promotes apoptosis, an activity attenuated by lymphoma-associated missense mutations [PMID:23842845]. In tumor settings, host STK10 supports anti-tumor cytotoxic T-cell activity and restrains tumor angiogenesis [PMID:36421382].","teleology":[{"year":1997,"claim":"Established the basic biochemical identity of STK10/LOK, answering whether it was a tyrosine or serine/threonine kinase and where it sat among kinase families.","evidence":"In vitro kinase assays on generic substrates and co-expression in COS7 cells","pmids":["9278426"],"confidence":"Medium","gaps":["No physiological substrate identified","No cellular function defined","MAP kinase pathways explicitly excluded but true effectors unknown"]},{"year":2000,"claim":"Connected STK10 to lymphocyte adhesion by showing it negatively regulates LFA-1 clustering, giving the kinase its first in vivo cellular role.","evidence":"LOK knockout mice with ICAM-1 binding and aggregation assays","pmids":["10692593"],"confidence":"Medium","gaps":["No molecular substrate linking the kinase to LFA-1 regulation","Mechanism of clustering control unknown"]},{"year":2003,"claim":"Identified STK10 as a candidate cell-cycle regulator acting upstream of PLK1, addressing whether the kinase had mitotic functions.","evidence":"Co-immunoprecipitation, in vitro kinase assay, and dominant-negative expression in NIH-3T3 cells","pmids":["12639966"],"confidence":"Medium","gaps":["PLK1 phosphosite not mapped","Physiological relevance of PLK1 activation in cycling cells not established"]},{"year":2009,"claim":"Defined STK10's central function as a major ERM kinase, answering what its key physiological substrate is and linking it to lymphocyte migration.","evidence":"In vitro peptide kinase assay with specificity profiling, MS localization, and LOK knockout mice","pmids":["19255442"],"confidence":"High","gaps":["Residual ERM phosphorylation indicates redundant kinases","How membrane enrichment restricts activity not yet defined"]},{"year":2012,"claim":"Showed how STK10 (with SLK) enforces spatial control of ERM activation, restricting ezrin phosphorylation to the apical/microvillar domain.","evidence":"RNAi, drug-resistant kinase rescue variants, and localization in polarized epithelial cells","pmids":["23209304"],"confidence":"High","gaps":["Molecular basis of local activation not resolved at this stage","Functional overlap between LOK and SLK not fully separated"]},{"year":2017,"claim":"Resolved the mechanistic basis for membrane-restricted ERM phosphorylation through a PIP2-dependent multi-step wedge mechanism.","evidence":"In vitro reconstitution, domain mapping, mutagenesis, and lipid-binding assays","pmids":["28430576"],"confidence":"High","gaps":["Structure of the kinase-ezrin complex not solved","Regulation of the kinase by upstream signals not addressed"]},{"year":2026,"claim":"Defined the structural and biophysical properties of the LOK C-terminal domain that target the kinase to ezrin at membranes.","evidence":"Lipid-binding and dimerization assays, MD simulations, and cell-based colocalization","pmids":["41958020"],"confidence":"Medium","gaps":["IBAR-like assignment based on prediction/simulation rather than experimental structure","Membrane curvature sensing not directly demonstrated"]},{"year":2013,"claim":"Linked STK10 to NF-κB suppression and apoptosis and to lymphoma-associated mutations, addressing a possible tumor-suppressive role.","evidence":"NF-κB reporter and dexamethasone apoptosis assays with site-directed PTCL mutants","pmids":["23842845"],"confidence":"Medium","gaps":["No direct biochemical target in the NF-κB pathway identified","Overexpression-based, kinase substrate unknown"]},{"year":2021,"claim":"Provided structural tools and chemical probes for STK10, addressing the lack of selective inhibitors and SLK/STK10 discrimination.","evidence":"X-ray crystallography of inhibitor complexes with cellular target engagement (extended by a 2025 macrocycle co-structure)","pmids":["34463505","41256983"],"confidence":"High","gaps":["Probes characterize binding, not in vivo pathway consequences","Selectivity beyond SLK not exhaustively profiled"]},{"year":2021,"claim":"Extended STK10's ERM and migration role to cancer cells, showing knockout alters migration/proliferation via p38 and phospho-ERM.","evidence":"CRISPR knockout in DU145 prostate cancer cells with phospho-signaling and phenotype readouts","pmids":["34149897"],"confidence":"Medium","gaps":["Mechanism linking STK10 to p38 not defined","Single cell line"]},{"year":2022,"claim":"Defined a host immune role for STK10 in anti-tumor immunity through cytotoxic T-cell activity and angiogenesis control.","evidence":"Stk10 knockout mice with tumor implantation and immune cell phenotyping","pmids":["36421382"],"confidence":"Medium","gaps":["Cell-intrinsic vs systemic contributions not separated","No substrate linking STK10 to CTL or vessel phenotypes"]},{"year":2026,"claim":"Identified a direct, physiologically validated STK10 substrate in platelets (ILK Ser343) and placed STK10 in hemostasis and arterial/venous thrombosis.","evidence":"Platelet-specific conditional KO, phosphoproteomics, IP-MS, in vitro kinase assay, and DVT/thrombosis models","pmids":["41055696","41791658"],"confidence":"High","gaps":["How ILK Ser343 phosphorylation drives downstream integrin/granule responses not fully resolved","Relationship between ERM and ILK substrate functions in platelets unclear"]},{"year":null,"claim":"How STK10's distinct activities — ERM phosphorylation, ILK phosphorylation, PLK1 activation, and NF-κB suppression — are integrated within single cell types and which upstream signals select among them remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model linking substrate choice to cellular context","Upstream activating signals defined only for platelet (Ca/PKC/PI3K) and ERK-tail inhibition contexts"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,4,8,14]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,4,14]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[8,15]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[4,5,8]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[5,17]}],"pathway":[{"term_id":"R-HSA-109582","term_label":"Hemostasis","supporting_discovery_ids":[14,16]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[1,11]}],"complexes":[],"partners":["EZR","MSN","PLK1","ILK"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O94804","full_name":"Serine/threonine-protein kinase 10","aliases":["Lymphocyte-oriented kinase"],"length_aa":968,"mass_kda":112.1,"function":"Serine/threonine-protein kinase involved in regulation of lymphocyte migration. Phosphorylates MSN, and possibly PLK1. Involved in regulation of lymphocyte migration by mediating phosphorylation of ERM proteins such as MSN. Acts as a negative regulator of MAP3K1/MEKK1. May also act as a cell cycle regulator by acting as a polo kinase kinase: mediates phosphorylation of PLK1 in vitro; however such data require additional evidences in vivo","subcellular_location":"Cell membrane","url":"https://www.uniprot.org/uniprotkb/O94804/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/STK10","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/STK10","total_profiled":1310},"omim":[{"mim_id":"616563","title":"STE20-LIKE PROTEIN KINASE; SLK","url":"https://www.omim.org/entry/616563"},{"mim_id":"603919","title":"SERINE/THREONINE PROTEIN KINASE 10; STK10","url":"https://www.omim.org/entry/603919"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Plasma membrane","reliability":"Supported"},{"location":"Nucleoplasm","reliability":"Additional"},{"location":"Nuclear bodies","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"bone marrow","ntpm":72.3}],"url":"https://www.proteinatlas.org/search/STK10"},"hgnc":{"alias_symbol":["LOK","PRO2729"],"prev_symbol":[]},"alphafold":{"accession":"O94804","domains":[{"cath_id":"3.30.200.20","chopping":"22-113","consensus_level":"medium","plddt":87.3238,"start":22,"end":113},{"cath_id":"1.10.510.10","chopping":"114-312","consensus_level":"medium","plddt":85.4528,"start":114,"end":312},{"cath_id":"-","chopping":"553-827","consensus_level":"medium","plddt":91.3703,"start":553,"end":827}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O94804","model_url":"https://alphafold.ebi.ac.uk/files/AF-O94804-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O94804-F1-predicted_aligned_error_v6.png","plddt_mean":73.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=STK10","jax_strain_url":"https://www.jax.org/strain/search?query=STK10"},"sequence":{"accession":"O94804","fasta_url":"https://rest.uniprot.org/uniprotkb/O94804.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O94804/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O94804"}},"corpus_meta":[{"pmid":"19255442","id":"PMC_19255442","title":"LOK is a major ERM kinase in resting lymphocytes and regulates cytoskeletal rearrangement through ERM phosphorylation.","date":"2009","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/19255442","citation_count":127,"is_preprint":false},{"pmid":"23209304","id":"PMC_23209304","title":"Local phosphocycling mediated by LOK/SLK restricts ezrin function to the apical aspect of epithelial cells.","date":"2012","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/23209304","citation_count":88,"is_preprint":false},{"pmid":"9278426","id":"PMC_9278426","title":"LOK is a novel mouse STE20-like protein kinase that is expressed predominantly in lymphocytes.","date":"1997","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9278426","citation_count":69,"is_preprint":false},{"pmid":"28430576","id":"PMC_28430576","title":"Ezrin activation by LOK phosphorylation involves a PIP2-dependent wedge mechanism.","date":"2017","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/28430576","citation_count":54,"is_preprint":false},{"pmid":"12639966","id":"PMC_12639966","title":"Stk10, a new member of the polo-like kinase kinase family highly expressed in hematopoietic tissue.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12639966","citation_count":52,"is_preprint":false},{"pmid":"27729408","id":"PMC_27729408","title":"DNA damage-induced Lok/CHK2 activation compromises germline stem cell self-renewal and lineage differentiation.","date":"2016","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/27729408","citation_count":33,"is_preprint":false},{"pmid":"10692593","id":"PMC_10692593","title":"Deficiency of a STE20/PAK family kinase LOK leads to the acceleration of LFA-1 clustering and cell adhesion of activated lymphocytes.","date":"2000","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/10692593","citation_count":27,"is_preprint":false},{"pmid":"11903060","id":"PMC_11903060","title":"Opposing roles of serine/threonine kinases MEKK1 and LOK in regulating the CD28 responsive element in T-cells.","date":"2002","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/11903060","citation_count":21,"is_preprint":false},{"pmid":"34149897","id":"PMC_34149897","title":"STK10 knockout inhibits cell migration and promotes cell proliferation via modulating the activity of ERM and p38 MAPK in prostate cancer cells.","date":"2021","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34149897","citation_count":19,"is_preprint":false},{"pmid":"10199912","id":"PMC_10199912","title":"Molecular cloning of the human gene STK10 encoding lymphocyte-oriented kinase, and comparative chromosomal mapping of the human, mouse, and rat homologues.","date":"1999","source":"Immunogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/10199912","citation_count":19,"is_preprint":false},{"pmid":"23869274","id":"PMC_23869274","title":"Migration of a Hem-o-Lok Clip to the Ureter Following Laparoscopic Partial Nephrectomy Presenting With Lower Urinary Tract Symptoms.","date":"2013","source":"International neurourology journal","url":"https://pubmed.ncbi.nlm.nih.gov/23869274","citation_count":19,"is_preprint":false},{"pmid":"23842845","id":"PMC_23842845","title":"STK10 missense mutations associated with anti-apoptotic function.","date":"2013","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/23842845","citation_count":18,"is_preprint":false},{"pmid":"31675459","id":"PMC_31675459","title":"Design and Analysis of the 4-Anilinoquin(az)oline Kinase Inhibition Profiles of GAK/SLK/STK10 Using Quantitative Structure-Activity Relationships.","date":"2019","source":"ChemMedChem","url":"https://pubmed.ncbi.nlm.nih.gov/31675459","citation_count":16,"is_preprint":false},{"pmid":"34463505","id":"PMC_34463505","title":"Discovery of a Potent Dual SLK/STK10 Inhibitor Based on a Maleimide Scaffold.","date":"2021","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/34463505","citation_count":10,"is_preprint":false},{"pmid":"37721438","id":"PMC_37721438","title":"VIRMA promotes neuron apoptosis via inducing m6A methylation of STK10 in spinal cord injury animal models.","date":"2023","source":"CNS neuroscience & therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/37721438","citation_count":7,"is_preprint":false},{"pmid":"35117313","id":"PMC_35117313","title":"Knockout of STK10 promotes the migration and invasion of cervical cancer cells.","date":"2020","source":"Translational cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/35117313","citation_count":5,"is_preprint":false},{"pmid":"32775668","id":"PMC_32775668","title":"Migration of Hem-O-Lok in Pelvicaliceal System Mimicking Renal Calculus Following Robotic Nephron-Sparing Surgery: A Case Report with Review of Literature.","date":"2020","source":"Journal of endourology case reports","url":"https://pubmed.ncbi.nlm.nih.gov/32775668","citation_count":5,"is_preprint":false},{"pmid":"41055696","id":"PMC_41055696","title":"STK10 regulates platelet function in arterial thrombosis and thromboinflammation.","date":"2026","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/41055696","citation_count":4,"is_preprint":false},{"pmid":"37923138","id":"PMC_37923138","title":"Kinome profiling identifies MARK3 and STK10 as potential therapeutic targets in uveal melanoma.","date":"2023","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/37923138","citation_count":4,"is_preprint":false},{"pmid":"36421382","id":"PMC_36421382","title":"Stk10 Deficiency in Mice Promotes Tumor Growth by Dysregulating the Tumor Microenvironment.","date":"2022","source":"Biology","url":"https://pubmed.ncbi.nlm.nih.gov/36421382","citation_count":3,"is_preprint":false},{"pmid":"28510197","id":"PMC_28510197","title":"Assimilates mobilization, stable canopy temperature and expression of expansin stabilizes grain weight in wheat cultivar LOK-1 under different soil moisture conditions.","date":"2017","source":"Botanical studies","url":"https://pubmed.ncbi.nlm.nih.gov/28510197","citation_count":3,"is_preprint":false},{"pmid":"29969952","id":"PMC_29969952","title":"Transmural migration of azygous vein Hem-O-lok clip causing food bolus 3 months following uneventful minimally invasive oesophagectomy.","date":"2018","source":"Acta chirurgica Belgica","url":"https://pubmed.ncbi.nlm.nih.gov/29969952","citation_count":3,"is_preprint":false},{"pmid":"21905501","id":"PMC_21905501","title":"Variations in the STK10 gene and possible associations with aspirin-intolerant asthma in a Korean population.","date":"2011","source":"Journal of investigational allergology & clinical immunology","url":"https://pubmed.ncbi.nlm.nih.gov/21905501","citation_count":2,"is_preprint":false},{"pmid":"41256983","id":"PMC_41256983","title":"Un-LOK-ing a New Approach for Conformational Selective Targeting of STK10 (LOK).","date":"2025","source":"ACS medicinal chemistry letters","url":"https://pubmed.ncbi.nlm.nih.gov/41256983","citation_count":1,"is_preprint":false},{"pmid":"27575462","id":"PMC_27575462","title":"The Application of Hem-O-Lok Clips Tied with Threads to Improve Surgical View in Retroperitoneal Laparoscopic Surgery for Renal Cell Carcinoma.","date":"2016","source":"Journal of laparoendoscopic & advanced surgical techniques. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/27575462","citation_count":1,"is_preprint":false},{"pmid":"21927701","id":"PMC_21927701","title":"A Feasible Technique for Transient Vascular Occlusion by Using a Vessel Loop and Hem-o-Lok Clips in Laparoscopic Partial Nephrectomy.","date":"2011","source":"Korean journal of urology","url":"https://pubmed.ncbi.nlm.nih.gov/21927701","citation_count":1,"is_preprint":false},{"pmid":"38761185","id":"PMC_38761185","title":"STK10 mutations block erythropoiesis in acquired pure red cell aplasia via impairing ribosome biogenesis.","date":"2024","source":"Annals of hematology","url":"https://pubmed.ncbi.nlm.nih.gov/38761185","citation_count":0,"is_preprint":false},{"pmid":"41791658","id":"PMC_41791658","title":"Deletion of platelet STK10 impairs deep vein thrombus formation.","date":"2026","source":"Journal of thrombosis and haemostasis : JTH","url":"https://pubmed.ncbi.nlm.nih.gov/41791658","citation_count":0,"is_preprint":false},{"pmid":"37967543","id":"PMC_37967543","title":"Transurethral Hem-o-lok Clip Ligation of the Distal Ureter in Retroperitoneal Laparoscopic Radical Nephroureterectomy.","date":"2023","source":"Urologia internationalis","url":"https://pubmed.ncbi.nlm.nih.gov/37967543","citation_count":0,"is_preprint":false},{"pmid":"41958020","id":"PMC_41958020","title":"The LOK C-terminus is an IBAR-like domain that facilitates membrane binding and ezrin colocalization.","date":"2026","source":"Biophysical journal","url":"https://pubmed.ncbi.nlm.nih.gov/41958020","citation_count":0,"is_preprint":false},{"pmid":"39511609","id":"PMC_39511609","title":"The clinical efficacy of monolayer suture combined with hem-o-lok clip in partial nephrectomy among patient with renal cell carcinoma: a quasi-experimental study.","date":"2024","source":"World journal of surgical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/39511609","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.01.19.25320804","title":"Systematic druggable genome-wide Mendelian randomization identifies therapeutic target genes for chronic periodontitis","date":"2025-01-20","source":"bioRxiv","url":"https://doi.org/10.1101/2025.01.19.25320804","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.11.15.688645","title":"Mechanism of ERK-mediated Rho Activation and Stress Fiber Assembly for Cell Migration","date":"2025-11-16","source":"bioRxiv","url":"https://doi.org/10.1101/2025.11.15.688645","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":17088,"output_tokens":4263,"usd":0.057605,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":12258,"output_tokens":3796,"usd":0.078095,"stage2_stop_reason":"end_turn"},"total_usd":0.1357,"stage1_batch_id":"msgbatch_013LWZZT6MWaqB9qyTSeQBmU","stage2_batch_id":"msgbatch_0194ZzQaTGNpruoNJBbEMbt3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1997,\n      \"finding\": \"LOK (STK10) is a serine/threonine kinase (not tyrosine kinase) that autophosphorylates and phosphorylates myelin basic protein and histone IIA; it belongs to the STE20 family with an N-terminal kinase domain and C-terminal coiled-coil/proline-rich region; it does not activate ERK, JNK, or p38 MAP kinases when co-expressed in COS7 cells.\",\n      \"method\": \"In vitro kinase assay, co-expression in COS7 cells, Western blot\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — in vitro kinase assay with substrate phosphorylation, single lab, foundational characterization paper\",\n      \"pmids\": [\"9278426\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"LOK-deficient mice show enhanced LFA-1 clustering and accelerated LFA-1/ICAM-mediated T cell aggregation upon mitogen stimulation, without changes in total LFA-1 or ICAM levels, indicating LOK negatively regulates LFA-1 clustering and lymphocyte adhesion.\",\n      \"method\": \"LOK knockout mouse model, soluble ICAM-1 binding assay, flow cytometry\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — genetic KO with defined cellular phenotype, single lab, single method\",\n      \"pmids\": [\"10692593\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Overexpression of LOK attenuates MEKK1-induced and Raji/SEE-induced CD28RE/AP1 reporter gene activation and IL-2 production in Jurkat T cells, indicating LOK opposes MEKK1 in the CD28 signaling pathway.\",\n      \"method\": \"Luciferase reporter assay, co-transfection in Jurkat cells, IL-2 measurement\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — overexpression reporter assay, single lab, no direct biochemical mechanism established\",\n      \"pmids\": [\"11903060\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"STK10 (human LOK) associates with PLK1 in cells and phosphorylates PLK1 in vitro; dominant-negative STK10 expression in NIH-3T3 cells causes increased DNA content, suggesting STK10 functions as a polo-like kinase kinase regulating PLK1.\",\n      \"method\": \"Co-immunoprecipitation, in vitro kinase assay, engineered NIH-3T3 cell lines with flow cytometry cell cycle analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP plus in vitro kinase assay plus dominant-negative cell phenotype, single lab\",\n      \"pmids\": [\"12639966\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"LOK is a major ERM kinase in lymphocytes: it is enriched at the plasma membrane near ERM proteins, directly phosphorylates moesin at its C-terminal threonine in vitro with preferential specificity (including unusual preference for Tyr at P-2), and LOK knockout mice show >50% reduction in ERM phosphorylation; loss of LOK enhances lymphocyte migration and polarization in response to chemokine.\",\n      \"method\": \"Mass spectrometry localization, immunofluorescence, in vitro peptide kinase assay, LOK kinase domain transfection, LOK knockout mouse model\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro kinase assay with substrate specificity profiling, genetic KO with defined phosphorylation and functional phenotype, multiple orthogonal methods\",\n      \"pmids\": [\"19255442\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"LOK and SLK are the relevant kinases driving apical restriction of ezrin in polarized epithelial cells; both kinases are enriched in microvilli and locally activated there; drug-resistant LOK/SLK variants are sufficient to restrict ezrin to the apical domain, while expression of their regulatory regions inhibits local ezrin phosphorylation by endogenous kinases.\",\n      \"method\": \"Proteomic approaches, RNAi knockdown, drug-resistant kinase variants, immunofluorescence localization\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNAi plus drug-resistant rescue variants plus localization studies, multiple orthogonal methods in one study\",\n      \"pmids\": [\"23209304\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Wild-type STK10 suppresses NF-κB activity and potentiates dexamethasone-induced apoptosis; PTCL-associated missense mutations (R634H, L85P, K277E) reduce this pro-apoptotic activity, with L85P and K277E having more profound anti-apoptotic effects than R634H.\",\n      \"method\": \"NF-κB reporter assay, apoptosis assay (dexamethasone), site-directed mutagenesis, transfection\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — functional assays with mutagenesis, single lab, no direct biochemical target of NF-κB regulation identified\",\n      \"pmids\": [\"23842845\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"In Drosophila GSCs, DNA damage activates Lok kinase, which is required for GSC loss and progeny differentiation defects; elimination of Lok or its kinase activity rescues these phenotypes; Lok-dependent signaling decreases expression of differentiation factor Bam.\",\n      \"method\": \"Genetic epistasis (lok knockout/kinase-dead), heat-shock I-CreI endonuclease and X-ray irradiation, immunofluorescence, Drosophila ovary model\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with multiple DNA-damage inducers and kinase-dead allele, Drosophila ortholog\",\n      \"pmids\": [\"27729408\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"LOK activates ezrin through a multi-step mechanism: (1) PIP2 binding to ezrin induces a conformational change; (2) the LOK C-terminal domain inserts to wedge apart the ezrin membrane- and F-actin-binding domains; (3) the LOK N-terminal kinase domain accesses a site 40 residues distal from the consensus sequence to phosphorylate the correct threonine. This ensures ezrin is only phosphorylated at the plasma membrane.\",\n      \"method\": \"In vitro reconstitution system, biochemical domain-mapping, mutagenesis, lipid-binding assays\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with mechanistic dissection, mutagenesis, and domain-mapping, multiple orthogonal biochemical methods in one study\",\n      \"pmids\": [\"28430576\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"STK10 knockout in prostate cancer DU145 cells inhibits cell migration and promotes proliferation; these effects are mediated via inhibition of p38 MAPK activation and reduced ERM protein phosphorylation.\",\n      \"method\": \"CRISPR-Cas9 knockout, Western blot (phospho-ERM, phospho-p38), migration and proliferation assays\",\n      \"journal\": \"Experimental and therapeutic medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — CRISPR KO with defined phospho-signaling readouts and cellular phenotypes, single lab\",\n      \"pmids\": [\"34149897\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Crystal structures of SLK and STK10 with maleimide-scaffold inhibitors were determined, revealing the binding mode and structural basis for selectivity between SLK and STK10; cellular target engagement assays confirmed inhibitor binding to STK10 in cells.\",\n      \"method\": \"X-ray crystallography, cellular target engagement assay (NanoBRET or similar), medicinal chemistry SAR\",\n      \"journal\": \"Journal of medicinal chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure with cellular validation, multiple inhibitor series with defined binding modes\",\n      \"pmids\": [\"34463505\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Host Stk10 knockout in mice results in increased tumor growth associated with decreased activated/effector cytotoxic T lymphocytes and increased vessel density in the tumor microenvironment, indicating STK10 modulates anti-tumor immunity through CTL activity and angiogenesis regulation.\",\n      \"method\": \"Stk10 knockout mouse model, tumor implantation assay, immunofluorescence/flow cytometry of tumor-infiltrating immune cells\",\n      \"journal\": \"Biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — genetic KO with in vivo tumor model and cellular immune phenotyping, single lab\",\n      \"pmids\": [\"36421382\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Knockdown of STK10 in K562 cells inhibits erythroid differentiation and promotes apoptosis, associated with inhibition of ribosome biogenesis, reduced ribosome levels, and activation of the p53 signaling pathway.\",\n      \"method\": \"shRNA knockdown in K562 cells, erythroid differentiation assay, ribosome profiling, Western blot (p53 pathway)\",\n      \"journal\": \"Annals of hematology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — knockdown with phenotype and pathway readouts, single lab, no direct biochemical mechanism linking STK10 to ribosome biogenesis\",\n      \"pmids\": [\"38761185\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"A co-crystal structure of a macrocyclic inhibitor bound to STK10 revealed a unique back-pocket binding mode distinct from SLK, providing structural basis for selective STK10 inhibition; compound 23 showed nanomolar STK10 activity in cells.\",\n      \"method\": \"X-ray co-crystallography, biophysical assays, cellular activity assays\",\n      \"journal\": \"ACS medicinal chemistry letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Weak — crystal structure with cellular validation, single study but direct structural evidence\",\n      \"pmids\": [\"41256983\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"STK10 in platelets is phosphorylated upon activation; platelet-specific STK10 knockout mice show impaired hemostasis, reduced platelet aggregation, α-granule release, αIIbβ3 activation, procoagulant activity, spreading, and clot retraction; STK10 directly phosphorylates integrin-linked kinase (ILK) at Ser343 as identified by immunoprecipitation-mass spectrometry and confirmed by in vitro phosphorylation assay; upstream, STK10 phosphorylation is regulated by calcium, PKC, and PI3K signaling.\",\n      \"method\": \"Platelet-specific conditional KO mice, quantitative phosphoproteomics, immunoprecipitation-mass spectrometry, in vitro kinase assay, platelet functional assays\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro kinase assay confirming direct ILK phosphorylation, conditional KO with multiple functional readouts, phosphoproteomics, multiple orthogonal methods\",\n      \"pmids\": [\"41055696\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"The LOK C-terminal domain (LOK-CTD) mediates colocalization of LOK with ezrin at the apical surface of epithelial cells; the LOK-CTD forms dimers, binds negatively charged phospholipids, and shares structural similarity to inverse BAR (IBAR) domains; these properties are required for LOK-ezrin colocalization.\",\n      \"method\": \"Biochemical assays (lipid binding, dimerization), predictive bioinformatics, molecular dynamics simulations (atomistic and coarse-grained), human cell-based colocalization assays\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical assays plus cell-based functional assays plus MD simulations, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"41958020\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Platelet STK10 deletion reduces deep vein thrombus formation in mice; STK10 phosphorylation and ILK (Ser343) phosphorylation are increased in platelets during DVT development; STK10 KO inhibits platelet-neutrophil interactions, NET formation, and platelet procoagulant activity in venous thrombi.\",\n      \"method\": \"Platelet-specific STK10 KO mice, inferior vena cava ligation DVT model, immunofluorescence, in vitro NET assay, phosphoprotein analysis\",\n      \"journal\": \"Journal of thrombosis and haemostasis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — conditional KO in vivo model with multiple cellular readouts, single lab, extends findings from companion arterial thrombosis paper\",\n      \"pmids\": [\"41791658\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ERK phosphorylates the C-terminal tail of LOK, inhibiting LOK's activation of Ezrin in the cell body; this releases Ezrin's inhibition of Rho (via ARHGAP18) and promotes stress fiber assembly and cell migration, placing LOK in an ERK→LOK→Ezrin→ARHGAP18→Rho pathway.\",\n      \"method\": \"Cell-based phosphorylation assays, genetic epistasis, Rho activity assays, stress fiber imaging, Ezrin activity assays\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — genetic epistasis and cell-based biochemical assays, preprint, single lab, not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.11.15.688645\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"STK10/LOK is a STE20-family serine/threonine kinase that primarily functions as a major ERM (ezrin/radixin/moesin) kinase enriched at the plasma membrane, where it phosphorylates ERM C-terminal threonines through a PIP2-dependent, multi-step wedge mechanism involving its C-terminal IBAR-like domain; it regulates lymphocyte migration, polarization, and LFA-1-mediated adhesion; acts as a PLK1-activating kinase; directly phosphorylates ILK at Ser343 in platelets to regulate arterial and venous thrombosis; suppresses NF-κB and promotes apoptosis; and is inhibited by ERK-mediated phosphorylation of its C-terminal tail to release Ezrin's inhibition of Rho for stress fiber-driven cell migration.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"STK10 (LOK) is a STE20-family serine/threonine kinase whose principal characterized function is acting as a major ERM (ezrin/radixin/moesin) kinase that couples plasma-membrane signaling to cortical actin organization, cell adhesion, and migration [#4, #5]. It autophosphorylates and uses an N-terminal kinase domain to directly phosphorylate the regulatory C-terminal threonine of ERM proteins with distinctive substrate specificity, and its loss reduces ERM phosphorylation by more than half in lymphocytes while enhancing chemokine-driven migration and polarization [#0, #4]. STK10 achieves spatial fidelity through a multi-step wedge mechanism in which PIP2 binding primes ezrin, the kinase's C-terminal domain inserts to separate the membrane- and actin-binding lobes, and the kinase domain then accesses a non-consensus phosphosite, ensuring phosphorylation occurs only at the membrane [#8]; the same C-terminal domain dimerizes, binds anionic phospholipids, resembles an inverse-BAR domain, and drives apical colocalization with ezrin in epithelial cells [#15, #5]. Beyond ERM regulation, STK10 negatively regulates LFA-1 clustering and lymphocyte adhesion [#1], associates with and phosphorylates PLK1 as a polo-like kinase kinase [#3], and in platelets is activated downstream of calcium/PKC/PI3K signaling to directly phosphorylate integrin-linked kinase (ILK) at Ser343, controlling aggregation, granule release, integrin activation, and both arterial-type and deep-vein thrombosis [#14, #16]. STK10 also suppresses NF-\\u03baB activity and promotes apoptosis, an activity attenuated by lymphoma-associated missense mutations [#6]. In tumor settings, host STK10 supports anti-tumor cytotoxic T-cell activity and restrains tumor angiogenesis [#11].\",\n  \"teleology\": [\n    {\n      \"year\": 1997,\n      \"claim\": \"Established the basic biochemical identity of STK10/LOK, answering whether it was a tyrosine or serine/threonine kinase and where it sat among kinase families.\",\n      \"evidence\": \"In vitro kinase assays on generic substrates and co-expression in COS7 cells\",\n      \"pmids\": [\"9278426\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No physiological substrate identified\", \"No cellular function defined\", \"MAP kinase pathways explicitly excluded but true effectors unknown\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Connected STK10 to lymphocyte adhesion by showing it negatively regulates LFA-1 clustering, giving the kinase its first in vivo cellular role.\",\n      \"evidence\": \"LOK knockout mice with ICAM-1 binding and aggregation assays\",\n      \"pmids\": [\"10692593\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No molecular substrate linking the kinase to LFA-1 regulation\", \"Mechanism of clustering control unknown\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Identified STK10 as a candidate cell-cycle regulator acting upstream of PLK1, addressing whether the kinase had mitotic functions.\",\n      \"evidence\": \"Co-immunoprecipitation, in vitro kinase assay, and dominant-negative expression in NIH-3T3 cells\",\n      \"pmids\": [\"12639966\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"PLK1 phosphosite not mapped\", \"Physiological relevance of PLK1 activation in cycling cells not established\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defined STK10's central function as a major ERM kinase, answering what its key physiological substrate is and linking it to lymphocyte migration.\",\n      \"evidence\": \"In vitro peptide kinase assay with specificity profiling, MS localization, and LOK knockout mice\",\n      \"pmids\": [\"19255442\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Residual ERM phosphorylation indicates redundant kinases\", \"How membrane enrichment restricts activity not yet defined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Showed how STK10 (with SLK) enforces spatial control of ERM activation, restricting ezrin phosphorylation to the apical/microvillar domain.\",\n      \"evidence\": \"RNAi, drug-resistant kinase rescue variants, and localization in polarized epithelial cells\",\n      \"pmids\": [\"23209304\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Molecular basis of local activation not resolved at this stage\", \"Functional overlap between LOK and SLK not fully separated\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Resolved the mechanistic basis for membrane-restricted ERM phosphorylation through a PIP2-dependent multi-step wedge mechanism.\",\n      \"evidence\": \"In vitro reconstitution, domain mapping, mutagenesis, and lipid-binding assays\",\n      \"pmids\": [\"28430576\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Structure of the kinase-ezrin complex not solved\", \"Regulation of the kinase by upstream signals not addressed\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Defined the structural and biophysical properties of the LOK C-terminal domain that target the kinase to ezrin at membranes.\",\n      \"evidence\": \"Lipid-binding and dimerization assays, MD simulations, and cell-based colocalization\",\n      \"pmids\": [\"41958020\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"IBAR-like assignment based on prediction/simulation rather than experimental structure\", \"Membrane curvature sensing not directly demonstrated\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Linked STK10 to NF-\\u03baB suppression and apoptosis and to lymphoma-associated mutations, addressing a possible tumor-suppressive role.\",\n      \"evidence\": \"NF-\\u03baB reporter and dexamethasone apoptosis assays with site-directed PTCL mutants\",\n      \"pmids\": [\"23842845\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No direct biochemical target in the NF-\\u03baB pathway identified\", \"Overexpression-based, kinase substrate unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Provided structural tools and chemical probes for STK10, addressing the lack of selective inhibitors and SLK/STK10 discrimination.\",\n      \"evidence\": \"X-ray crystallography of inhibitor complexes with cellular target engagement (extended by a 2025 macrocycle co-structure)\",\n      \"pmids\": [\"34463505\", \"41256983\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Probes characterize binding, not in vivo pathway consequences\", \"Selectivity beyond SLK not exhaustively profiled\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extended STK10's ERM and migration role to cancer cells, showing knockout alters migration/proliferation via p38 and phospho-ERM.\",\n      \"evidence\": \"CRISPR knockout in DU145 prostate cancer cells with phospho-signaling and phenotype readouts\",\n      \"pmids\": [\"34149897\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Mechanism linking STK10 to p38 not defined\", \"Single cell line\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined a host immune role for STK10 in anti-tumor immunity through cytotoxic T-cell activity and angiogenesis control.\",\n      \"evidence\": \"Stk10 knockout mice with tumor implantation and immune cell phenotyping\",\n      \"pmids\": [\"36421382\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Cell-intrinsic vs systemic contributions not separated\", \"No substrate linking STK10 to CTL or vessel phenotypes\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Identified a direct, physiologically validated STK10 substrate in platelets (ILK Ser343) and placed STK10 in hemostasis and arterial/venous thrombosis.\",\n      \"evidence\": \"Platelet-specific conditional KO, phosphoproteomics, IP-MS, in vitro kinase assay, and DVT/thrombosis models\",\n      \"pmids\": [\"41055696\", \"41791658\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"How ILK Ser343 phosphorylation drives downstream integrin/granule responses not fully resolved\", \"Relationship between ERM and ILK substrate functions in platelets unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How STK10's distinct activities — ERM phosphorylation, ILK phosphorylation, PLK1 activation, and NF-\\u03baB suppression — are integrated within single cell types and which upstream signals select among them remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No unified model linking substrate choice to cellular context\", \"Upstream activating signals defined only for platelet (Ca/PKC/PI3K) and ERK-tail inhibition contexts\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 4, 8, 14]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 4, 14]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [8, 15]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [4, 5, 8]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [5, 17]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-109582\", \"supporting_discovery_ids\": [14, 16]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [1, 11]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"EZR\", \"MSN\", \"PLK1\", \"ILK\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}