{"gene":"ICMT","run_date":"2026-04-28T18:06:53","timeline":{"discoveries":[{"year":2011,"finding":"Crystal structure of a prokaryotic ICMT ortholog revealed the enzyme comprises a core of five transmembrane α-helices with a cofactor (SAM)-binding pocket in a highly conserved C-terminal catalytic subdomain, and a tunnel linking the reactive methyl group of SAM to the inner membrane provides access for the prenyl lipid substrate, explaining how the integral membrane methyltransferase accommodates both a hydrophilic cofactor and a lipophilic prenyl substrate.","method":"X-ray crystallography of prokaryotic ICMT ortholog","journal":"Molecular Cell","confidence":"High","confidence_rationale":"Tier 1 — crystal structure with mechanistic interpretation of active site architecture","pmids":["22195972"],"is_preprint":false},{"year":2018,"finding":"X-ray crystal structure of eukaryotic ICMT at 2.3 Å resolution (in complex with SAM cofactor, an ordered lipid, and a monobody inhibitor) demonstrated that the active site spans cytosolic and membrane-exposed regions, indicating distinct entry routes for the cytosolic methyl donor SAM and for prenylcysteine substrates associated with the ER membrane.","method":"X-ray crystallography with monobody inhibitor co-crystallization","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 — high-resolution eukaryotic structure with bound cofactor and inhibitor, published in Nature","pmids":["29342140"],"is_preprint":false},{"year":2004,"finding":"Human ICMT catalysis proceeds through an ordered sequential kinetic mechanism in which SAM binds first; dead-end competitive inhibitor analysis with S-farnesylthioacetic acid and product inhibition studies with S-adenosyl-L-homocysteine (AdoHcy) and N-acetyl-S-farnesyl-L-cysteine methylester established that AdoHcy is the final product released.","method":"In vitro kinetic analysis with recombinant human ICMT, dead-end inhibitor and product inhibition studies","journal":"BMC Biochemistry","confidence":"High","confidence_rationale":"Tier 1 — rigorous in vitro kinetic mechanism established with multiple inhibitor analyses","pmids":["15625008"],"is_preprint":false},{"year":2014,"finding":"Scanning mutagenesis of nearly half of the residues of the Anopheles gambiae ICMT ortholog, combined with methyltransferase assays, identified 62 mutations causing reduced or undetectable activity; residues within transmembrane regions contribute to the isoprenylcysteine binding site, and certain substitutions caused substrate inhibition, while cytosolic residues contribute to AdoMet binding in a manner similar to a related prokaryotic methyltransferase (Ma MTase).","method":"Scanning mutagenesis combined with in vitro methyltransferase activity assays","journal":"Journal of Biological Chemistry","confidence":"High","confidence_rationale":"Tier 1 — systematic mutagenesis across nearly half the protein with functional readout","pmids":["25059662"],"is_preprint":false},{"year":2004,"finding":"Genetic inactivation of Icmt in mouse embryonic fibroblasts expressing oncogenic K-Ras inhibited K-Ras-induced oncogenic transformation (soft agar and nude mice assays); mechanistically, Icmt loss caused accelerated RhoA protein turnover/reduced RhoA levels, a Ras/Erk1/2-dependent increase in p21(Cip1), and deletion of p21(Cip1) rescued soft-agar growth, placing Icmt upstream of RhoA stability and p21(Cip1) induction.","method":"Conditional Cre-mediated Icmt knockout in K-Ras-expressing fibroblasts, soft agar assay, nude mouse xenograft, p21(Cip1) double knockout rescue","journal":"Journal of Clinical Investigation","confidence":"High","confidence_rationale":"Tier 2 — genetic epistasis with multiple orthogonal readouts, replicated in vitro and in vivo","pmids":["14966563"],"is_preprint":false},{"year":2006,"finding":"ICMT is the only enzyme in mammalian cells capable of methylating isoprenylated CAAX proteins; in addition to CAAX proteins (including Ras), ICMT also methylates the CXC class of isoprenylated Rab proteins. In the absence of carboxyl methylation, Ras proteins are mislocalized away from the plasma membrane and exhibit a shift in electrophoretic mobility.","method":"Genetic knockout studies and subcellular fractionation/localization in Icmt-deficient cells","journal":"Methods in Enzymology","confidence":"High","confidence_rationale":"Tier 2 — genetic loss-of-function with direct localization readout, consistent with multiple independent studies","pmids":["16757321"],"is_preprint":false},{"year":2021,"finding":"Among the four RAS proteins, NRAS uniquely requires ICMT for delivery to the plasma membrane because it has only a single palmitoylation site; ICMT-mediated carboxyl methylation is required for efficient NRAS palmitoylation, for binding to the prenyl-protein chaperone PDE6δ, and for homeostatic Golgi flux, all of which direct NRAS to the plasma membrane.","method":"ICMT knockout cells, photoactivation and FRAP of GFP-NRAS, palmitoylation assays, PDE6δ binding assays, subcellular localization imaging","journal":"Life Science Alliance","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (FRAP, palmitoylation assay, co-IP with chaperone) in ICMT-null cells","pmids":["33579760"],"is_preprint":false},{"year":2016,"finding":"Retina-specific Icmt knockout in mice caused progressive loss of rod and cone photoreceptor function; ICMT-mediated methylation is required for proper membrane association of isoprenylated transducin and cone phosphodiesterase 6 (PDE6α'), and for maintaining protein levels of transducin, PDE6α', and cone GRK1, demonstrating that carboxyl methylation is essential for phototransduction protein membrane targeting.","method":"Retina-specific conditional Icmt knockout mice, electroretinography, subcellular fractionation, immunoblotting","journal":"Journal of Neuroscience","confidence":"High","confidence_rationale":"Tier 2 — tissue-specific KO with defined molecular phenotype and multiple protein readouts","pmids":["27147662"],"is_preprint":false},{"year":2021,"finding":"Genetic knockout of Icmt in HGPS mice improved survival and restored vascular smooth muscle cell numbers in the aorta; pharmacologic ICMT inhibition with compound C75 delayed senescence and stimulated proliferation of late-passage HGPS cells and Zmpste24-deficient fibroblasts, acting specifically through ICMT (C75 had no effect on Zmpste24-deficient cells also lacking Icmt), implicating ICMT-mediated methylation of farnesylated progerin in HGPS pathology.","method":"Icmt knockout HGPS mouse model, ICMT inhibitor C75, proliferation and senescence assays, specificity confirmed in double-KO cells","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 — genetic and pharmacologic evidence with on-target specificity controls, in vivo and in vitro","pmids":["33526168"],"is_preprint":false},{"year":2019,"finding":"Pharmacologic ICMT inhibition with compound UCM-1336 (IC50 = 2 μM) significantly impaired membrane association of all four Ras isoforms, decreased Ras-GTP levels, and inhibited downstream Ras signaling pathways, demonstrating that ICMT-mediated methylation is required for Ras membrane localization and activity regardless of activating mutation.","method":"ICMT inhibitor treatment, subcellular fractionation, Ras activation (pull-down) assay, downstream signaling by immunoblot, in vivo AML model","journal":"Journal of Medicinal Chemistry","confidence":"High","confidence_rationale":"Tier 2 — selective inhibitor with direct Ras localization and activity readouts plus in vivo validation","pmids":["31181882"],"is_preprint":false},{"year":2007,"finding":"Chemical inhibition of ICMT with AGGC caused redistribution and aggregation of the ER chaperone GRP94, followed by GRP94 protein degradation; these effects were dependent on RhoA activity and caspase activation, and GRP94 depletion augmented ICMT-inhibition-induced endothelial cell apoptosis, linking ICMT/RhoA signaling to the unfolded protein response.","method":"ICMT inhibitor (AGGC) treatment, 2D-PAGE proteomics, immunofluorescence, overexpression of constitutively active RhoA, caspase inhibition","journal":"American Journal of Respiratory Cell and Molecular Biology","confidence":"Medium","confidence_rationale":"Tier 2 — multiple methods but single lab; GRP94 link is correlative with some functional rescue","pmids":["17347446"],"is_preprint":false},{"year":2014,"finding":"miR-100 directly inhibits ICMT expression by binding to the ICMT 3'-UTR; ICMT knockdown phenocopied miR-100 overexpression in suppressing lamellipodia formation and MMP2 activation, and constitutively active Rac1(Q61L) rescued the anti-metastatic effect of miR-100, placing ICMT upstream of Rac1 in a signaling axis controlling HCC cell migration and invasion.","method":"Luciferase 3'-UTR reporter assay, gain- and loss-of-function studies, Rac1(Q61L) rescue, mouse orthotopic xenograft","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 — direct 3'-UTR validation plus epistasis rescue, single lab","pmids":["25361001"],"is_preprint":false},{"year":2019,"finding":"Wild-type p53 represses ICMT transcription by binding to the ICMT promoter region between -209 and -14, as established by ICMT-luciferase reporters, promoter truncation analysis, and ChIP assays; in contrast, cancer-associated p53 mutants positively regulate ICMT expression through a distinct promoter region.","method":"ICMT-luciferase reporter assay, promoter truncation analysis, chromatin immunoprecipitation (ChIP)","journal":"Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP plus reporter assay establish direct p53 recruitment to ICMT promoter, single lab","pmids":["30655292"],"is_preprint":false},{"year":2022,"finding":"PFKFB4 physically interacts with ICMT (shown by co-immunoprecipitation), promotes the ICMT/RAS interaction, controls RAS localization at the plasma membrane, and activates AKT signaling to enhance melanoma cell migration, revealing a glycolysis-independent function of PFKFB4 acting through ICMT.","method":"Co-immunoprecipitation, RAS localization imaging, AKT signaling assays, migration assays","journal":"Life Science Alliance","confidence":"Medium","confidence_rationale":"Tier 3 — single Co-IP plus functional readouts, single lab","pmids":["35914811"],"is_preprint":false},{"year":2024,"finding":"ICMT overexpression enhanced invadopodia formation and promoted lung metastasis in vivo; ICMT overexpression promoted migration and invasion in vitro, acting through its substrates that regulate actin cytoskeleton dynamics.","method":"ICMT overexpression, invadopodia formation assay, in vivo lung metastasis model, migration and invasion assays","journal":"Biochimie","confidence":"Medium","confidence_rationale":"Tier 2 — defined cellular phenotype (invadopodia) with in vivo validation, single lab","pmids":["38301884"],"is_preprint":false},{"year":2011,"finding":"Prenylcysteine-based ICMT inhibitors (analogs 1a and 1b) exhibit mixed-mode inhibition of human ICMT with the competitive component predominating; cellular treatment with inhibitor 1b alters subcellular localization of GFP-KRas and inhibits Ras activation and Erk phosphorylation in Jurkat cells.","method":"Kinetic inhibitor analysis (Ki determination), GFP-KRas localization microscopy, Ras activation assay, immunoblotting","journal":"Bioorganic & Medicinal Chemistry","confidence":"Medium","confidence_rationale":"Tier 1-2 — in vitro kinetic mechanism plus cellular localization and signaling readouts","pmids":["22142613"],"is_preprint":false},{"year":2025,"finding":"RAC1P29S requires C-terminal prenylation (and thus ICMT-mediated methylation) to drive MAPKi resistance in BRAF(V600E) melanoma; mechanistically, combined ICMT inhibition (cysmethynil) and MAPK pathway inhibition impaired nuclear translocation of TAZ, whose transcriptional activity accounts for MAPKi resistance in RAC1P29S cells, and constitutively active TAZ phenocopied RAC1P29S-driven resistance.","method":"RAC1P29S/C189S prenylation-site mutant, cysmethynil ICMT inhibitor, TAZ nuclear translocation assay, constitutively active TAZ rescue, in vitro and in vivo tumor models","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 — genetic and pharmacologic epistasis with defined downstream mechanism (TAZ), preprint not yet peer-reviewed","pmids":["bio_10.1101_2025.08.10.669587"],"is_preprint":true}],"current_model":"ICMT is an endoplasmic reticulum-resident integral membrane methyltransferase with a five-transmembrane-helix core whose active site spans cytosolic and membrane-embedded regions, catalyzing transfer of the methyl group from SAM (which binds first in an ordered sequential mechanism) through a hydrophobic tunnel to the carboxylate of the isoprenylcysteine at the C-terminus of CAAX and CXC proteins (including all four RAS isoforms, RHO GTPases, and RAB proteins), thereby promoting their plasma membrane targeting, palmitoylation, chaperone (PDE6δ) binding, and downstream signaling; its expression is transcriptionally repressed by wild-type p53 and its loss or inhibition mislocalizes RAS, destabilizes RhoA, induces p21(Cip1)-dependent growth arrest, and blocks oncogenic transformation."},"narrative":{"teleology":[{"year":2004,"claim":"Establishing the catalytic mechanism: the question of substrate binding order and cofactor release was resolved, showing ICMT uses an ordered sequential mechanism with SAM binding first and S-adenosyl-L-homocysteine released last, providing a kinetic framework for inhibitor design.","evidence":"In vitro kinetic analysis with recombinant human ICMT using dead-end competitive inhibitors and product inhibition studies","pmids":["15625008"],"confidence":"High","gaps":["No structural basis for the ordered mechanism at this point","Kinetics determined with small-molecule substrates, not full-length prenylated proteins"]},{"year":2004,"claim":"Defining the biological requirement for ICMT in oncogenic signaling: genetic inactivation of Icmt in K-Ras-transformed fibroblasts blocked transformation, revealing that ICMT loss destabilizes RhoA and induces p21(Cip1)-dependent growth arrest—establishing ICMT as a gatekeeper for RAS-driven oncogenesis.","evidence":"Conditional Cre-mediated Icmt knockout in K-Ras-expressing MEFs, soft agar colony formation, nude mouse xenograft, p21 double-knockout rescue","pmids":["14966563"],"confidence":"High","gaps":["Whether ICMT loss blocks transformation by all RAS isoforms equally was untested","Mechanism linking RhoA destabilization to p21 induction was not fully resolved"]},{"year":2006,"claim":"Establishing ICMT as the sole mammalian isoprenylcysteine carboxyl methyltransferase: ICMT methylates both CAAX and CXC (Rab) classes of prenylated proteins, and its absence mislocalizes RAS from the plasma membrane.","evidence":"Icmt-knockout cells with subcellular fractionation and electrophoretic mobility shift of unmethylated Ras","pmids":["16757321"],"confidence":"High","gaps":["Relative dependence of individual Ras isoforms on methylation for membrane targeting was not differentiated"]},{"year":2011,"claim":"Revealing the structural basis of membrane-embedded catalysis: the first crystal structure of an ICMT ortholog showed a five-transmembrane-helix core with a tunnel connecting the SAM-binding pocket to the membrane interior, explaining how a single enzyme accommodates both hydrophilic and lipophilic substrates.","evidence":"X-ray crystallography of prokaryotic ICMT ortholog","pmids":["22195972"],"confidence":"High","gaps":["Prokaryotic ortholog may not capture all features of mammalian ICMT","Substrate-bound structure not obtained"]},{"year":2014,"claim":"Mapping the functional residue landscape: systematic mutagenesis identified 62 activity-reducing mutations, confirming that transmembrane residues form the isoprenylcysteine binding site and cytosolic residues engage SAM, and revealed that certain substitutions cause substrate inhibition.","evidence":"Scanning mutagenesis of ~50% of Anopheles gambiae ICMT residues with methyltransferase activity assays","pmids":["25059662"],"confidence":"High","gaps":["Insect ortholog mutagenesis; not all findings may transfer quantitatively to human ICMT","No crystal structure of mutant enzymes to confirm structural interpretation"]},{"year":2016,"claim":"Demonstrating a physiological role for ICMT in sensory neurobiology: retina-specific Icmt knockout caused progressive photoreceptor degeneration by disrupting membrane association and stability of prenylated phototransduction proteins including transducin and cone PDE6.","evidence":"Retina-specific conditional Icmt knockout mice with electroretinography and subcellular fractionation","pmids":["27147662"],"confidence":"High","gaps":["Whether methylation-deficient transducin retains partial signaling activity was not determined","Contribution of individual ICMT substrates to the degenerative phenotype was not dissected"]},{"year":2018,"claim":"Achieving the eukaryotic structure: the 2.3 Å crystal structure of ICMT in complex with SAM and a monobody inhibitor confirmed distinct cytosolic and membrane-embedded entry routes for cofactor and substrate, providing a template for rational drug design.","evidence":"X-ray crystallography of eukaryotic ICMT with monobody co-crystallization","pmids":["29342140"],"confidence":"High","gaps":["No structure with a bound prenylcysteine substrate or full-length protein substrate","Dynamics of substrate entry through the membrane not captured by crystallography"]},{"year":2019,"claim":"Pharmacologic validation of ICMT as a druggable target for RAS-driven cancers: ICMT inhibitor UCM-1336 impaired membrane association and GTP loading of all four RAS isoforms and inhibited downstream signaling, with efficacy in an AML model.","evidence":"UCM-1336 treatment with subcellular fractionation, Ras-GTP pull-down, immunoblotting, and in vivo AML xenograft","pmids":["31181882"],"confidence":"High","gaps":["Off-target effects on non-RAS prenylated substrates not fully characterized","Long-term resistance mechanisms to ICMT inhibition not explored"]},{"year":2019,"claim":"Identifying transcriptional control of ICMT: wild-type p53 directly represses ICMT transcription by binding its promoter, while cancer-associated p53 mutants upregulate ICMT, connecting tumor suppressor status to the prenylation-methylation axis.","evidence":"ICMT-luciferase reporter, promoter truncation, and chromatin immunoprecipitation (ChIP) for p53","pmids":["30655292"],"confidence":"Medium","gaps":["Physiological relevance of p53-dependent ICMT repression in tumor suppression not demonstrated in vivo","Mechanism by which mutant p53 activates ICMT expression not resolved"]},{"year":2021,"claim":"Resolving isoform-specific dependence on methylation: NRAS uniquely requires ICMT for plasma membrane delivery because its single palmitoylation site makes it dependent on methylation for efficient palmitoylation and PDE6δ chaperone binding.","evidence":"ICMT-knockout cells with photoactivation/FRAP of GFP-NRAS, palmitoylation assays, and PDE6δ co-immunoprecipitation","pmids":["33579760"],"confidence":"High","gaps":["Whether NRAS-specific dependence translates to differential therapeutic vulnerability in NRAS-mutant cancers was not tested","Quantitative contribution of each step (palmitoylation, PDE6δ binding, Golgi flux) not resolved"]},{"year":2021,"claim":"Establishing ICMT as a therapeutic target in progeria: genetic Icmt deletion and pharmacologic ICMT inhibition (C75) improved survival in HGPS mice and rescued senescence in HGPS cells, with on-target specificity confirmed in double-knockout controls.","evidence":"Icmt knockout in HGPS mouse model, C75 inhibitor treatment, proliferation/senescence assays, Icmt/Zmpste24 double-KO specificity control","pmids":["33526168"],"confidence":"High","gaps":["Whether ICMT inhibition reverses established disease or only prevents progression was not determined","Potential toxicity from global ICMT inhibition in vivo not thoroughly assessed"]},{"year":null,"claim":"A substrate-bound structure of eukaryotic ICMT with a prenylated peptide or full-length protein substrate has not been determined, leaving the precise geometry of methyl transfer and the structural basis for substrate selectivity among diverse prenylated proteins unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No substrate-bound eukaryotic ICMT structure","Structural basis for selectivity among CAAX vs CXC substrates unknown","Allosteric regulation and role of interactors such as PFKFB4 structurally uncharacterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,1,2,3,5]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[1,5,6]}],"pathway":[],"complexes":[],"partners":["KRAS","NRAS","RHOA","RAC1","PDE6D","PFKFB4"],"other_free_text":[]},"mechanistic_narrative":"ICMT is an endoplasmic reticulum-resident integral membrane methyltransferase that catalyzes the terminal post-translational modification of isoprenylated CAAX and CXC proteins—including all four RAS isoforms, RHO GTPases, and RAB proteins—by transferring a methyl group from S-adenosylmethionine (SAM) to the C-terminal isoprenylcysteine carboxylate, thereby promoting plasma membrane targeting and downstream signaling [PMID:16757321, PMID:31181882]. Structurally, ICMT contains a five-transmembrane-helix core with a hydrophobic tunnel linking the cytosolic SAM-binding pocket to the membrane-embedded prenylcysteine-binding site, and catalysis proceeds through an ordered sequential mechanism in which SAM binds first [PMID:22195972, PMID:29342140, PMID:15625008]. Loss of ICMT mislocalizes RAS from the plasma membrane, destabilizes RhoA, induces p21(Cip1)-dependent growth arrest, and blocks oncogenic K-Ras-driven transformation, while its expression is transcriptionally repressed by wild-type p53 [PMID:14966563, PMID:30655292]. Genetic deletion of Icmt in a Hutchinson-Gilford progeria syndrome mouse model improves survival, implicating ICMT-mediated methylation of farnesylated progerin in disease pathology [PMID:33526168]."},"prefetch_data":{"uniprot":{"accession":"O60725","full_name":"Protein-S-isoprenylcysteine O-methyltransferase","aliases":["Isoprenylcysteine carboxylmethyltransferase","Prenylated protein carboxyl methyltransferase","PPMT","Prenylcysteine carboxyl methyltransferase","pcCMT"],"length_aa":284,"mass_kda":31.9,"function":"Catalyzes the post-translational methylation of isoprenylated C-terminal cysteine residues","subcellular_location":"Endoplasmic reticulum membrane","url":"https://www.uniprot.org/uniprotkb/O60725/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ICMT","classification":"Not Classified","n_dependent_lines":162,"n_total_lines":1208,"dependency_fraction":0.13410596026490065},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CCDC47","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/ICMT","total_profiled":1310},"omim":[{"mim_id":"606480","title":"ZINC METALLOPROTEINASE STE24; ZMPSTE24","url":"https://www.omim.org/entry/606480"},{"mim_id":"605851","title":"ISOPRENYLCYSTEINE CARBOXYLMETHYLTRANSFERASE; ICMT","url":"https://www.omim.org/entry/605851"},{"mim_id":"150330","title":"LAMIN A/C; LMNA","url":"https://www.omim.org/entry/150330"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ICMT"},"hgnc":{"alias_symbol":["PCCMT","HSTE14","PPMT"],"prev_symbol":[]},"alphafold":{"accession":"O60725","domains":[{"cath_id":"-","chopping":"12-128","consensus_level":"medium","plddt":93.1671,"start":12,"end":128},{"cath_id":"1.20.120.1630","chopping":"129-284","consensus_level":"medium","plddt":95.7662,"start":129,"end":284}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O60725","model_url":"https://alphafold.ebi.ac.uk/files/AF-O60725-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O60725-F1-predicted_aligned_error_v6.png","plddt_mean":93.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ICMT","jax_strain_url":"https://www.jax.org/strain/search?query=ICMT"},"sequence":{"accession":"O60725","fasta_url":"https://rest.uniprot.org/uniprotkb/O60725.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O60725/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O60725"}},"corpus_meta":[{"pmid":"14966563","id":"PMC_14966563","title":"Inactivation of Icmt inhibits transformation by oncogenic K-Ras and B-Raf.","date":"2004","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/14966563","citation_count":133,"is_preprint":false},{"pmid":"23949910","id":"PMC_23949910","title":"18F-ICMT-11, a caspase-3-specific PET tracer for apoptosis: biodistribution and radiation dosimetry.","date":"2013","source":"Journal of nuclear medicine : official publication, Society of Nuclear Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/23949910","citation_count":74,"is_preprint":false},{"pmid":"11748165","id":"PMC_11748165","title":"icmT is essential for pore formation-mediated egress of Legionella pneumophila from mammalian and protozoan cells.","date":"2002","source":"Infection and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/11748165","citation_count":70,"is_preprint":false},{"pmid":"22195972","id":"PMC_22195972","title":"Mechanism of isoprenylcysteine carboxyl methylation from the crystal structure of the integral membrane methyltransferase ICMT.","date":"2011","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/22195972","citation_count":45,"is_preprint":false},{"pmid":"25361001","id":"PMC_25361001","title":"Downregulation of microRNA-100 enhances the ICMT-Rac1 signaling and promotes metastasis of hepatocellular carcinoma cells.","date":"2014","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/25361001","citation_count":39,"is_preprint":false},{"pmid":"21661760","id":"PMC_21661760","title":"Discovery and SAR of methylated tetrahydropyranyl derivatives as inhibitors of isoprenylcysteine carboxyl methyltransferase (ICMT).","date":"2011","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21661760","citation_count":37,"is_preprint":false},{"pmid":"29342140","id":"PMC_29342140","title":"Atomic structure of the eukaryotic intramembrane RAS methyltransferase ICMT.","date":"2018","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/29342140","citation_count":34,"is_preprint":false},{"pmid":"31181882","id":"PMC_31181882","title":"A Potent Isoprenylcysteine Carboxylmethyltransferase (ICMT) Inhibitor Improves Survival in Ras-Driven Acute Myeloid Leukemia.","date":"2019","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/31181882","citation_count":34,"is_preprint":false},{"pmid":"30259091","id":"PMC_30259091","title":"Clinical translation of [18F]ICMT-11 for measuring chemotherapy-induced caspase 3/7 activation in breast and lung cancer.","date":"2018","source":"European journal of nuclear medicine and molecular imaging","url":"https://pubmed.ncbi.nlm.nih.gov/30259091","citation_count":32,"is_preprint":false},{"pmid":"11918802","id":"PMC_11918802","title":"The C-terminus of IcmT is essential for pore formation and for intracellular trafficking of Legionella pneumophila within Acanthamoeba polyphaga.","date":"2002","source":"Molecular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/11918802","citation_count":31,"is_preprint":false},{"pmid":"24618809","id":"PMC_24618809","title":"Preclinical assessment of carboplatin treatment efficacy in lung cancer by 18F-ICMT-11-positron emission tomography.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24618809","citation_count":28,"is_preprint":false},{"pmid":"15625008","id":"PMC_15625008","title":"Analysis of the kinetic mechanism of recombinant human isoprenylcysteine carboxylmethyltransferase (Icmt).","date":"2004","source":"BMC biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/15625008","citation_count":27,"is_preprint":false},{"pmid":"23514631","id":"PMC_23514631","title":"Functionalized indoleamines as potent, drug-like inhibitors of isoprenylcysteine carboxyl methyltransferase (Icmt).","date":"2013","source":"European journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/23514631","citation_count":23,"is_preprint":false},{"pmid":"33526168","id":"PMC_33526168","title":"A small-molecule ICMT inhibitor delays senescence of Hutchinson-Gilford progeria syndrome cells.","date":"2021","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/33526168","citation_count":21,"is_preprint":false},{"pmid":"31451223","id":"PMC_31451223","title":"ICMT contributes to hepatocellular carcinoma growth, survival, migration and chemoresistance via multiple oncogenic pathways.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/31451223","citation_count":16,"is_preprint":false},{"pmid":"30655292","id":"PMC_30655292","title":"Isoprenylcysteine carboxy methyltransferase (ICMT) is associated with tumor aggressiveness and its expression is controlled by the p53 tumor suppressor.","date":"2019","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30655292","citation_count":16,"is_preprint":false},{"pmid":"17347446","id":"PMC_17347446","title":"Inhibition of ICMT induces endothelial cell apoptosis through GRP94.","date":"2007","source":"American journal of respiratory cell and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/17347446","citation_count":15,"is_preprint":false},{"pmid":"36339587","id":"PMC_36339587","title":"Licoricidin combats gastric cancer by targeting the ICMT/Ras pathway in vitro and in vivo.","date":"2022","source":"Frontiers in pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/36339587","citation_count":14,"is_preprint":false},{"pmid":"22142613","id":"PMC_22142613","title":"Amide-modified prenylcysteine based Icmt inhibitors: Structure-activity relationships, kinetic analysis and cellular characterization.","date":"2011","source":"Bioorganic & medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22142613","citation_count":14,"is_preprint":false},{"pmid":"15621435","id":"PMC_15621435","title":"Structure-function analysis of the C-terminus of IcmT of Legionella pneumophila in pore formation-mediated egress from macrophages.","date":"2005","source":"FEMS microbiology letters","url":"https://pubmed.ncbi.nlm.nih.gov/15621435","citation_count":12,"is_preprint":false},{"pmid":"26959120","id":"PMC_26959120","title":"Icmt inhibition exerts anti-angiogenic and anti-hyperpermeability activities impeding malignant pleural effusion.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26959120","citation_count":11,"is_preprint":false},{"pmid":"35914811","id":"PMC_35914811","title":"PFKFB4 interacts with ICMT and activates RAS/AKT signaling-dependent cell migration in melanoma.","date":"2022","source":"Life science alliance","url":"https://pubmed.ncbi.nlm.nih.gov/35914811","citation_count":10,"is_preprint":false},{"pmid":"16757321","id":"PMC_16757321","title":"Genetic and pharmacologic analyses of the role of Icmt in Ras membrane association and function.","date":"2006","source":"Methods in enzymology","url":"https://pubmed.ncbi.nlm.nih.gov/16757321","citation_count":10,"is_preprint":false},{"pmid":"35312113","id":"PMC_35312113","title":"CircRNA hsa_circ_0018289 exerts an oncogenic role in cervical cancer progression through miR-1294/ICMT axis.","date":"2022","source":"Journal of clinical laboratory analysis","url":"https://pubmed.ncbi.nlm.nih.gov/35312113","citation_count":10,"is_preprint":false},{"pmid":"25059662","id":"PMC_25059662","title":"Mutational analysis of the integral membrane methyltransferase isoprenylcysteine carboxyl methyltransferase (ICMT) reveals potential substrate binding sites.","date":"2014","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25059662","citation_count":9,"is_preprint":false},{"pmid":"33579760","id":"PMC_33579760","title":"NRAS is unique among RAS proteins in requiring ICMT for trafficking to the plasma membrane.","date":"2021","source":"Life science alliance","url":"https://pubmed.ncbi.nlm.nih.gov/33579760","citation_count":8,"is_preprint":false},{"pmid":"27147662","id":"PMC_27147662","title":"Deficiency of Isoprenylcysteine Carboxyl Methyltransferase (ICMT) Leads to Progressive Loss of Photoreceptor Function.","date":"2016","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/27147662","citation_count":8,"is_preprint":false},{"pmid":"34984187","id":"PMC_34984187","title":"Lidocaine exerts anticancer activity in bladder cancer by targeting isoprenylcysteine carboxylmethyltransferase (ICMT).","date":"2021","source":"Translational andrology and urology","url":"https://pubmed.ncbi.nlm.nih.gov/34984187","citation_count":8,"is_preprint":false},{"pmid":"32781531","id":"PMC_32781531","title":"Development of [18F]ICMT-11 for Imaging Caspase-3/7 Activity during Therapy-Induced Apoptosis.","date":"2020","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/32781531","citation_count":7,"is_preprint":false},{"pmid":"26718043","id":"PMC_26718043","title":"9 Structure and function of isoprenylcysteine carboxylmethyltransferase (Icmt): A key enzyme in CaaX processing.","date":"2007","source":"The Enzymes","url":"https://pubmed.ncbi.nlm.nih.gov/26718043","citation_count":6,"is_preprint":false},{"pmid":"29794225","id":"PMC_29794225","title":"Depicting Changes in Tumor Biology in Response to Cetuximab Monotherapy or Combination Therapy by Apoptosis and Proliferation Imaging Using 18F-ICMT-11 and 18F-FLT PET.","date":"2018","source":"Journal of nuclear medicine : official publication, Society of Nuclear Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29794225","citation_count":6,"is_preprint":false},{"pmid":"23725486","id":"PMC_23725486","title":"Mapping peptide thiol accessibility in membranes using a quaternary ammonium isotope-coded mass tag (ICMT).","date":"2013","source":"Bioconjugate chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/23725486","citation_count":5,"is_preprint":false},{"pmid":"30125103","id":"PMC_30125103","title":"Use of an Isotope-Coded Mass Tag (ICMT) Method To Determine the Orientation of Cholesterol Oxidase on Model Membranes.","date":"2018","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30125103","citation_count":3,"is_preprint":false},{"pmid":"38301884","id":"PMC_38301884","title":"Isoprenylcysteine carboxyl methyltransferase (ICMT) promotes invadopodia formation and metastasis in cancer cells.","date":"2024","source":"Biochimie","url":"https://pubmed.ncbi.nlm.nih.gov/38301884","citation_count":2,"is_preprint":false},{"pmid":"38835128","id":"PMC_38835128","title":"In silico Exploration of a Novel ICMT Inhibitor with More Solubility than Cysmethynil against Membrane Localization of KRAS Mutant in Colorectal Cancer.","date":"2024","source":"Current computer-aided drug design","url":"https://pubmed.ncbi.nlm.nih.gov/38835128","citation_count":1,"is_preprint":false},{"pmid":"41808116","id":"PMC_41808116","title":"ICMT deficiency ameliorates weight loss and mortality, but not tumor formation in a mouse model of liver cancer.","date":"2026","source":"Cell communication and signaling : CCS","url":"https://pubmed.ncbi.nlm.nih.gov/41808116","citation_count":0,"is_preprint":false},{"pmid":"41877819","id":"PMC_41877819","title":"Targeting ICMT: A promising strategy in cancer treatment (Review).","date":"2026","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/41877819","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.08.10.669587","title":"Concurrent inhibition of ICMT and RAF/MEK suppresses RAC1P29S-driven MAPKi resistance in BRAFV600E melanoma by regulating TAZ activity","date":"2025-08-12","source":"bioRxiv","url":"https://doi.org/10.1101/2025.08.10.669587","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":21201,"output_tokens":4206,"usd":0.063347},"stage2":{"model":"claude-opus-4-6","input_tokens":7667,"output_tokens":5642,"usd":0.269078},"total_usd":0.332425,"stage1_batch_id":"msgbatch_01DzrsgVgYngY1hvM3DU1yF2","stage2_batch_id":"msgbatch_01KA9b7iRzKFdhoBUT7vCvSe","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2011,\n      \"finding\": \"Crystal structure of a prokaryotic ICMT ortholog revealed the enzyme comprises a core of five transmembrane α-helices with a cofactor (SAM)-binding pocket in a highly conserved C-terminal catalytic subdomain, and a tunnel linking the reactive methyl group of SAM to the inner membrane provides access for the prenyl lipid substrate, explaining how the integral membrane methyltransferase accommodates both a hydrophilic cofactor and a lipophilic prenyl substrate.\",\n      \"method\": \"X-ray crystallography of prokaryotic ICMT ortholog\",\n      \"journal\": \"Molecular Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure with mechanistic interpretation of active site architecture\",\n      \"pmids\": [\"22195972\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"X-ray crystal structure of eukaryotic ICMT at 2.3 Å resolution (in complex with SAM cofactor, an ordered lipid, and a monobody inhibitor) demonstrated that the active site spans cytosolic and membrane-exposed regions, indicating distinct entry routes for the cytosolic methyl donor SAM and for prenylcysteine substrates associated with the ER membrane.\",\n      \"method\": \"X-ray crystallography with monobody inhibitor co-crystallization\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — high-resolution eukaryotic structure with bound cofactor and inhibitor, published in Nature\",\n      \"pmids\": [\"29342140\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Human ICMT catalysis proceeds through an ordered sequential kinetic mechanism in which SAM binds first; dead-end competitive inhibitor analysis with S-farnesylthioacetic acid and product inhibition studies with S-adenosyl-L-homocysteine (AdoHcy) and N-acetyl-S-farnesyl-L-cysteine methylester established that AdoHcy is the final product released.\",\n      \"method\": \"In vitro kinetic analysis with recombinant human ICMT, dead-end inhibitor and product inhibition studies\",\n      \"journal\": \"BMC Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — rigorous in vitro kinetic mechanism established with multiple inhibitor analyses\",\n      \"pmids\": [\"15625008\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Scanning mutagenesis of nearly half of the residues of the Anopheles gambiae ICMT ortholog, combined with methyltransferase assays, identified 62 mutations causing reduced or undetectable activity; residues within transmembrane regions contribute to the isoprenylcysteine binding site, and certain substitutions caused substrate inhibition, while cytosolic residues contribute to AdoMet binding in a manner similar to a related prokaryotic methyltransferase (Ma MTase).\",\n      \"method\": \"Scanning mutagenesis combined with in vitro methyltransferase activity assays\",\n      \"journal\": \"Journal of Biological Chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — systematic mutagenesis across nearly half the protein with functional readout\",\n      \"pmids\": [\"25059662\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Genetic inactivation of Icmt in mouse embryonic fibroblasts expressing oncogenic K-Ras inhibited K-Ras-induced oncogenic transformation (soft agar and nude mice assays); mechanistically, Icmt loss caused accelerated RhoA protein turnover/reduced RhoA levels, a Ras/Erk1/2-dependent increase in p21(Cip1), and deletion of p21(Cip1) rescued soft-agar growth, placing Icmt upstream of RhoA stability and p21(Cip1) induction.\",\n      \"method\": \"Conditional Cre-mediated Icmt knockout in K-Ras-expressing fibroblasts, soft agar assay, nude mouse xenograft, p21(Cip1) double knockout rescue\",\n      \"journal\": \"Journal of Clinical Investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis with multiple orthogonal readouts, replicated in vitro and in vivo\",\n      \"pmids\": [\"14966563\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"ICMT is the only enzyme in mammalian cells capable of methylating isoprenylated CAAX proteins; in addition to CAAX proteins (including Ras), ICMT also methylates the CXC class of isoprenylated Rab proteins. In the absence of carboxyl methylation, Ras proteins are mislocalized away from the plasma membrane and exhibit a shift in electrophoretic mobility.\",\n      \"method\": \"Genetic knockout studies and subcellular fractionation/localization in Icmt-deficient cells\",\n      \"journal\": \"Methods in Enzymology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic loss-of-function with direct localization readout, consistent with multiple independent studies\",\n      \"pmids\": [\"16757321\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Among the four RAS proteins, NRAS uniquely requires ICMT for delivery to the plasma membrane because it has only a single palmitoylation site; ICMT-mediated carboxyl methylation is required for efficient NRAS palmitoylation, for binding to the prenyl-protein chaperone PDE6δ, and for homeostatic Golgi flux, all of which direct NRAS to the plasma membrane.\",\n      \"method\": \"ICMT knockout cells, photoactivation and FRAP of GFP-NRAS, palmitoylation assays, PDE6δ binding assays, subcellular localization imaging\",\n      \"journal\": \"Life Science Alliance\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (FRAP, palmitoylation assay, co-IP with chaperone) in ICMT-null cells\",\n      \"pmids\": [\"33579760\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Retina-specific Icmt knockout in mice caused progressive loss of rod and cone photoreceptor function; ICMT-mediated methylation is required for proper membrane association of isoprenylated transducin and cone phosphodiesterase 6 (PDE6α'), and for maintaining protein levels of transducin, PDE6α', and cone GRK1, demonstrating that carboxyl methylation is essential for phototransduction protein membrane targeting.\",\n      \"method\": \"Retina-specific conditional Icmt knockout mice, electroretinography, subcellular fractionation, immunoblotting\",\n      \"journal\": \"Journal of Neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — tissue-specific KO with defined molecular phenotype and multiple protein readouts\",\n      \"pmids\": [\"27147662\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Genetic knockout of Icmt in HGPS mice improved survival and restored vascular smooth muscle cell numbers in the aorta; pharmacologic ICMT inhibition with compound C75 delayed senescence and stimulated proliferation of late-passage HGPS cells and Zmpste24-deficient fibroblasts, acting specifically through ICMT (C75 had no effect on Zmpste24-deficient cells also lacking Icmt), implicating ICMT-mediated methylation of farnesylated progerin in HGPS pathology.\",\n      \"method\": \"Icmt knockout HGPS mouse model, ICMT inhibitor C75, proliferation and senescence assays, specificity confirmed in double-KO cells\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic and pharmacologic evidence with on-target specificity controls, in vivo and in vitro\",\n      \"pmids\": [\"33526168\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Pharmacologic ICMT inhibition with compound UCM-1336 (IC50 = 2 μM) significantly impaired membrane association of all four Ras isoforms, decreased Ras-GTP levels, and inhibited downstream Ras signaling pathways, demonstrating that ICMT-mediated methylation is required for Ras membrane localization and activity regardless of activating mutation.\",\n      \"method\": \"ICMT inhibitor treatment, subcellular fractionation, Ras activation (pull-down) assay, downstream signaling by immunoblot, in vivo AML model\",\n      \"journal\": \"Journal of Medicinal Chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — selective inhibitor with direct Ras localization and activity readouts plus in vivo validation\",\n      \"pmids\": [\"31181882\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Chemical inhibition of ICMT with AGGC caused redistribution and aggregation of the ER chaperone GRP94, followed by GRP94 protein degradation; these effects were dependent on RhoA activity and caspase activation, and GRP94 depletion augmented ICMT-inhibition-induced endothelial cell apoptosis, linking ICMT/RhoA signaling to the unfolded protein response.\",\n      \"method\": \"ICMT inhibitor (AGGC) treatment, 2D-PAGE proteomics, immunofluorescence, overexpression of constitutively active RhoA, caspase inhibition\",\n      \"journal\": \"American Journal of Respiratory Cell and Molecular Biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple methods but single lab; GRP94 link is correlative with some functional rescue\",\n      \"pmids\": [\"17347446\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"miR-100 directly inhibits ICMT expression by binding to the ICMT 3'-UTR; ICMT knockdown phenocopied miR-100 overexpression in suppressing lamellipodia formation and MMP2 activation, and constitutively active Rac1(Q61L) rescued the anti-metastatic effect of miR-100, placing ICMT upstream of Rac1 in a signaling axis controlling HCC cell migration and invasion.\",\n      \"method\": \"Luciferase 3'-UTR reporter assay, gain- and loss-of-function studies, Rac1(Q61L) rescue, mouse orthotopic xenograft\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct 3'-UTR validation plus epistasis rescue, single lab\",\n      \"pmids\": [\"25361001\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Wild-type p53 represses ICMT transcription by binding to the ICMT promoter region between -209 and -14, as established by ICMT-luciferase reporters, promoter truncation analysis, and ChIP assays; in contrast, cancer-associated p53 mutants positively regulate ICMT expression through a distinct promoter region.\",\n      \"method\": \"ICMT-luciferase reporter assay, promoter truncation analysis, chromatin immunoprecipitation (ChIP)\",\n      \"journal\": \"Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus reporter assay establish direct p53 recruitment to ICMT promoter, single lab\",\n      \"pmids\": [\"30655292\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"PFKFB4 physically interacts with ICMT (shown by co-immunoprecipitation), promotes the ICMT/RAS interaction, controls RAS localization at the plasma membrane, and activates AKT signaling to enhance melanoma cell migration, revealing a glycolysis-independent function of PFKFB4 acting through ICMT.\",\n      \"method\": \"Co-immunoprecipitation, RAS localization imaging, AKT signaling assays, migration assays\",\n      \"journal\": \"Life Science Alliance\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single Co-IP plus functional readouts, single lab\",\n      \"pmids\": [\"35914811\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ICMT overexpression enhanced invadopodia formation and promoted lung metastasis in vivo; ICMT overexpression promoted migration and invasion in vitro, acting through its substrates that regulate actin cytoskeleton dynamics.\",\n      \"method\": \"ICMT overexpression, invadopodia formation assay, in vivo lung metastasis model, migration and invasion assays\",\n      \"journal\": \"Biochimie\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — defined cellular phenotype (invadopodia) with in vivo validation, single lab\",\n      \"pmids\": [\"38301884\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Prenylcysteine-based ICMT inhibitors (analogs 1a and 1b) exhibit mixed-mode inhibition of human ICMT with the competitive component predominating; cellular treatment with inhibitor 1b alters subcellular localization of GFP-KRas and inhibits Ras activation and Erk phosphorylation in Jurkat cells.\",\n      \"method\": \"Kinetic inhibitor analysis (Ki determination), GFP-KRas localization microscopy, Ras activation assay, immunoblotting\",\n      \"journal\": \"Bioorganic & Medicinal Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro kinetic mechanism plus cellular localization and signaling readouts\",\n      \"pmids\": [\"22142613\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RAC1P29S requires C-terminal prenylation (and thus ICMT-mediated methylation) to drive MAPKi resistance in BRAF(V600E) melanoma; mechanistically, combined ICMT inhibition (cysmethynil) and MAPK pathway inhibition impaired nuclear translocation of TAZ, whose transcriptional activity accounts for MAPKi resistance in RAC1P29S cells, and constitutively active TAZ phenocopied RAC1P29S-driven resistance.\",\n      \"method\": \"RAC1P29S/C189S prenylation-site mutant, cysmethynil ICMT inhibitor, TAZ nuclear translocation assay, constitutively active TAZ rescue, in vitro and in vivo tumor models\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic and pharmacologic epistasis with defined downstream mechanism (TAZ), preprint not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.08.10.669587\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"ICMT is an endoplasmic reticulum-resident integral membrane methyltransferase with a five-transmembrane-helix core whose active site spans cytosolic and membrane-embedded regions, catalyzing transfer of the methyl group from SAM (which binds first in an ordered sequential mechanism) through a hydrophobic tunnel to the carboxylate of the isoprenylcysteine at the C-terminus of CAAX and CXC proteins (including all four RAS isoforms, RHO GTPases, and RAB proteins), thereby promoting their plasma membrane targeting, palmitoylation, chaperone (PDE6δ) binding, and downstream signaling; its expression is transcriptionally repressed by wild-type p53 and its loss or inhibition mislocalizes RAS, destabilizes RhoA, induces p21(Cip1)-dependent growth arrest, and blocks oncogenic transformation.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"ICMT is an endoplasmic reticulum-resident integral membrane methyltransferase that catalyzes the terminal post-translational modification of isoprenylated CAAX and CXC proteins—including all four RAS isoforms, RHO GTPases, and RAB proteins—by transferring a methyl group from S-adenosylmethionine (SAM) to the C-terminal isoprenylcysteine carboxylate, thereby promoting plasma membrane targeting and downstream signaling [PMID:16757321, PMID:31181882]. Structurally, ICMT contains a five-transmembrane-helix core with a hydrophobic tunnel linking the cytosolic SAM-binding pocket to the membrane-embedded prenylcysteine-binding site, and catalysis proceeds through an ordered sequential mechanism in which SAM binds first [PMID:22195972, PMID:29342140, PMID:15625008]. Loss of ICMT mislocalizes RAS from the plasma membrane, destabilizes RhoA, induces p21(Cip1)-dependent growth arrest, and blocks oncogenic K-Ras-driven transformation, while its expression is transcriptionally repressed by wild-type p53 [PMID:14966563, PMID:30655292]. Genetic deletion of Icmt in a Hutchinson-Gilford progeria syndrome mouse model improves survival, implicating ICMT-mediated methylation of farnesylated progerin in disease pathology [PMID:33526168].\",\n  \"teleology\": [\n    {\n      \"year\": 2004,\n      \"claim\": \"Establishing the catalytic mechanism: the question of substrate binding order and cofactor release was resolved, showing ICMT uses an ordered sequential mechanism with SAM binding first and S-adenosyl-L-homocysteine released last, providing a kinetic framework for inhibitor design.\",\n      \"evidence\": \"In vitro kinetic analysis with recombinant human ICMT using dead-end competitive inhibitors and product inhibition studies\",\n      \"pmids\": [\"15625008\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structural basis for the ordered mechanism at this point\", \"Kinetics determined with small-molecule substrates, not full-length prenylated proteins\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Defining the biological requirement for ICMT in oncogenic signaling: genetic inactivation of Icmt in K-Ras-transformed fibroblasts blocked transformation, revealing that ICMT loss destabilizes RhoA and induces p21(Cip1)-dependent growth arrest—establishing ICMT as a gatekeeper for RAS-driven oncogenesis.\",\n      \"evidence\": \"Conditional Cre-mediated Icmt knockout in K-Ras-expressing MEFs, soft agar colony formation, nude mouse xenograft, p21 double-knockout rescue\",\n      \"pmids\": [\"14966563\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether ICMT loss blocks transformation by all RAS isoforms equally was untested\", \"Mechanism linking RhoA destabilization to p21 induction was not fully resolved\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Establishing ICMT as the sole mammalian isoprenylcysteine carboxyl methyltransferase: ICMT methylates both CAAX and CXC (Rab) classes of prenylated proteins, and its absence mislocalizes RAS from the plasma membrane.\",\n      \"evidence\": \"Icmt-knockout cells with subcellular fractionation and electrophoretic mobility shift of unmethylated Ras\",\n      \"pmids\": [\"16757321\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative dependence of individual Ras isoforms on methylation for membrane targeting was not differentiated\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Revealing the structural basis of membrane-embedded catalysis: the first crystal structure of an ICMT ortholog showed a five-transmembrane-helix core with a tunnel connecting the SAM-binding pocket to the membrane interior, explaining how a single enzyme accommodates both hydrophilic and lipophilic substrates.\",\n      \"evidence\": \"X-ray crystallography of prokaryotic ICMT ortholog\",\n      \"pmids\": [\"22195972\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Prokaryotic ortholog may not capture all features of mammalian ICMT\", \"Substrate-bound structure not obtained\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Mapping the functional residue landscape: systematic mutagenesis identified 62 activity-reducing mutations, confirming that transmembrane residues form the isoprenylcysteine binding site and cytosolic residues engage SAM, and revealed that certain substitutions cause substrate inhibition.\",\n      \"evidence\": \"Scanning mutagenesis of ~50% of Anopheles gambiae ICMT residues with methyltransferase activity assays\",\n      \"pmids\": [\"25059662\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Insect ortholog mutagenesis; not all findings may transfer quantitatively to human ICMT\", \"No crystal structure of mutant enzymes to confirm structural interpretation\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrating a physiological role for ICMT in sensory neurobiology: retina-specific Icmt knockout caused progressive photoreceptor degeneration by disrupting membrane association and stability of prenylated phototransduction proteins including transducin and cone PDE6.\",\n      \"evidence\": \"Retina-specific conditional Icmt knockout mice with electroretinography and subcellular fractionation\",\n      \"pmids\": [\"27147662\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether methylation-deficient transducin retains partial signaling activity was not determined\", \"Contribution of individual ICMT substrates to the degenerative phenotype was not dissected\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Achieving the eukaryotic structure: the 2.3 Å crystal structure of ICMT in complex with SAM and a monobody inhibitor confirmed distinct cytosolic and membrane-embedded entry routes for cofactor and substrate, providing a template for rational drug design.\",\n      \"evidence\": \"X-ray crystallography of eukaryotic ICMT with monobody co-crystallization\",\n      \"pmids\": [\"29342140\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structure with a bound prenylcysteine substrate or full-length protein substrate\", \"Dynamics of substrate entry through the membrane not captured by crystallography\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Pharmacologic validation of ICMT as a druggable target for RAS-driven cancers: ICMT inhibitor UCM-1336 impaired membrane association and GTP loading of all four RAS isoforms and inhibited downstream signaling, with efficacy in an AML model.\",\n      \"evidence\": \"UCM-1336 treatment with subcellular fractionation, Ras-GTP pull-down, immunoblotting, and in vivo AML xenograft\",\n      \"pmids\": [\"31181882\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Off-target effects on non-RAS prenylated substrates not fully characterized\", \"Long-term resistance mechanisms to ICMT inhibition not explored\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identifying transcriptional control of ICMT: wild-type p53 directly represses ICMT transcription by binding its promoter, while cancer-associated p53 mutants upregulate ICMT, connecting tumor suppressor status to the prenylation-methylation axis.\",\n      \"evidence\": \"ICMT-luciferase reporter, promoter truncation, and chromatin immunoprecipitation (ChIP) for p53\",\n      \"pmids\": [\"30655292\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological relevance of p53-dependent ICMT repression in tumor suppression not demonstrated in vivo\", \"Mechanism by which mutant p53 activates ICMT expression not resolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Resolving isoform-specific dependence on methylation: NRAS uniquely requires ICMT for plasma membrane delivery because its single palmitoylation site makes it dependent on methylation for efficient palmitoylation and PDE6δ chaperone binding.\",\n      \"evidence\": \"ICMT-knockout cells with photoactivation/FRAP of GFP-NRAS, palmitoylation assays, and PDE6δ co-immunoprecipitation\",\n      \"pmids\": [\"33579760\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether NRAS-specific dependence translates to differential therapeutic vulnerability in NRAS-mutant cancers was not tested\", \"Quantitative contribution of each step (palmitoylation, PDE6δ binding, Golgi flux) not resolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Establishing ICMT as a therapeutic target in progeria: genetic Icmt deletion and pharmacologic ICMT inhibition (C75) improved survival in HGPS mice and rescued senescence in HGPS cells, with on-target specificity confirmed in double-knockout controls.\",\n      \"evidence\": \"Icmt knockout in HGPS mouse model, C75 inhibitor treatment, proliferation/senescence assays, Icmt/Zmpste24 double-KO specificity control\",\n      \"pmids\": [\"33526168\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether ICMT inhibition reverses established disease or only prevents progression was not determined\", \"Potential toxicity from global ICMT inhibition in vivo not thoroughly assessed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A substrate-bound structure of eukaryotic ICMT with a prenylated peptide or full-length protein substrate has not been determined, leaving the precise geometry of methyl transfer and the structural basis for substrate selectivity among diverse prenylated proteins unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No substrate-bound eukaryotic ICMT structure\", \"Structural basis for selectivity among CAAX vs CXC substrates unknown\", \"Allosteric regulation and role of interactors such as PFKFB4 structurally uncharacterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 1, 2, 3, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [1, 5, 6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": []}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"KRAS\", \"NRAS\", \"RHOA\", \"RAC1\", \"PDE6D\", \"PFKFB4\"],\n    \"other_free_text\": []\n  }\n}\n```\n\nWait, I need to fix the pathway field — it should use Reactome term_ids, not GO. Let me reconsider.\n\n```json\n{\n  \"mechanistic_narrative\": \"ICMT is an endoplasmic reticulum-resident integral membrane methyltransferase that catalyzes the terminal post-translational modification of isoprenylated CAAX and CXC proteins—including all four RAS isoforms, RHO GTPases, and RAB proteins—by transferring a methyl group from S-adenosylmethionine (SAM) to the C-terminal isoprenylcysteine carboxylate, thereby promoting plasma membrane targeting and downstream signaling [PMID:16757321, PMID:31181882]. Structurally, ICMT contains a five-transmembrane-helix core with a hydrophobic tunnel linking the cytosolic SAM-binding pocket to the membrane-embedded prenylcysteine-binding site, and catalysis proceeds through an ordered sequential mechanism in which SAM binds first [PMID:22195972, PMID:29342140, PMID:15625008]. Loss of ICMT mislocalizes RAS from the plasma membrane, destabilizes RhoA, induces p21(Cip1)-dependent growth arrest, and blocks oncogenic K-Ras-driven transformation, while its expression is transcriptionally repressed by wild-type p53 [PMID:14966563, PMID:30655292]. Genetic deletion of Icmt in a Hutchinson-Gilford progeria syndrome mouse model improves survival, implicating ICMT-mediated methylation of farnesylated progerin in disease pathology [PMID:33526168].\",\n  \"teleology\": [\n    {\n      \"year\": 2004,\n      \"claim\": \"Establishing the catalytic mechanism: the question of substrate binding order and cofactor release was resolved, showing ICMT uses an ordered sequential mechanism with SAM binding first and S-adenosyl-L-homocysteine released last, providing a kinetic framework for inhibitor design.\",\n      \"evidence\": \"In vitro kinetic analysis with recombinant human ICMT using dead-end competitive inhibitors and product inhibition studies\",\n      \"pmids\": [\"15625008\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structural basis for the ordered mechanism at this point\", \"Kinetics determined with small-molecule substrates, not full-length prenylated proteins\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Defining the biological requirement for ICMT in oncogenic signaling: genetic inactivation of Icmt in K-Ras-transformed fibroblasts blocked transformation, revealing that ICMT loss destabilizes RhoA and induces p21(Cip1)-dependent growth arrest—establishing ICMT as a gatekeeper for RAS-driven oncogenesis.\",\n      \"evidence\": \"Conditional Cre-mediated Icmt knockout in K-Ras-expressing MEFs, soft agar colony formation, nude mouse xenograft, p21 double-knockout rescue\",\n      \"pmids\": [\"14966563\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether ICMT loss blocks transformation by all RAS isoforms equally was untested\", \"Mechanism linking RhoA destabilization to p21 induction was not fully resolved\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Establishing ICMT as the sole mammalian isoprenylcysteine carboxyl methyltransferase: ICMT methylates both CAAX and CXC (Rab) classes of prenylated proteins, and its absence mislocalizes RAS from the plasma membrane.\",\n      \"evidence\": \"Icmt-knockout cells with subcellular fractionation and electrophoretic mobility shift of unmethylated Ras\",\n      \"pmids\": [\"16757321\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative dependence of individual Ras isoforms on methylation for membrane targeting was not differentiated\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Revealing the structural basis of membrane-embedded catalysis: the first crystal structure of an ICMT ortholog showed a five-transmembrane-helix core with a tunnel connecting the SAM-binding pocket to the membrane interior, explaining how a single enzyme accommodates both hydrophilic and lipophilic substrates.\",\n      \"evidence\": \"X-ray crystallography of prokaryotic ICMT ortholog\",\n      \"pmids\": [\"22195972\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Prokaryotic ortholog may not capture all features of mammalian ICMT\", \"Substrate-bound structure not obtained\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Mapping the functional residue landscape: systematic mutagenesis identified 62 activity-reducing mutations, confirming that transmembrane residues form the isoprenylcysteine binding site and cytosolic residues engage SAM, and revealed that certain substitutions cause substrate inhibition.\",\n      \"evidence\": \"Scanning mutagenesis of ~50% of Anopheles gambiae ICMT residues with methyltransferase activity assays\",\n      \"pmids\": [\"25059662\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Insect ortholog mutagenesis; not all findings may transfer quantitatively to human ICMT\", \"No crystal structure of mutant enzymes to confirm structural interpretation\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrating a physiological role for ICMT in sensory neurobiology: retina-specific Icmt knockout caused progressive photoreceptor degeneration by disrupting membrane association and stability of prenylated phototransduction proteins including transducin and cone PDE6.\",\n      \"evidence\": \"Retina-specific conditional Icmt knockout mice with electroretinography and subcellular fractionation\",\n      \"pmids\": [\"27147662\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether methylation-deficient transducin retains partial signaling activity was not determined\", \"Contribution of individual ICMT substrates to the degenerative phenotype was not dissected\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Achieving the eukaryotic structure: the 2.3 Å crystal structure of ICMT in complex with SAM and a monobody inhibitor confirmed distinct cytosolic and membrane-embedded entry routes for cofactor and substrate, providing a template for rational drug design.\",\n      \"evidence\": \"X-ray crystallography of eukaryotic ICMT with monobody co-crystallization\",\n      \"pmids\": [\"29342140\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structure with a bound prenylcysteine substrate or full-length protein substrate\", \"Dynamics of substrate entry through the membrane not captured by crystallography\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Pharmacologic validation of ICMT as a druggable target for RAS-driven cancers: ICMT inhibitor UCM-1336 impaired membrane association and GTP loading of all four RAS isoforms and inhibited downstream signaling, with efficacy in an AML model.\",\n      \"evidence\": \"UCM-1336 treatment with subcellular fractionation, Ras-GTP pull-down, immunoblotting, and in vivo AML xenograft\",\n      \"pmids\": [\"31181882\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Off-target effects on non-RAS prenylated substrates not fully characterized\", \"Long-term resistance mechanisms to ICMT inhibition not explored\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identifying transcriptional control of ICMT: wild-type p53 directly represses ICMT transcription by binding its promoter, while cancer-associated p53 mutants upregulate ICMT, connecting tumor suppressor status to the prenylation-methylation axis.\",\n      \"evidence\": \"ICMT-luciferase reporter, promoter truncation, and chromatin immunoprecipitation (ChIP) for p53\",\n      \"pmids\": [\"30655292\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological relevance of p53-dependent ICMT repression in tumor suppression not demonstrated in vivo\", \"Mechanism by which mutant p53 activates ICMT expression not resolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Resolving isoform-specific dependence on methylation: NRAS uniquely requires ICMT for plasma membrane delivery because its single palmitoylation site makes it dependent on methylation for efficient palmitoylation and PDE6δ chaperone binding.\",\n      \"evidence\": \"ICMT-knockout cells with photoactivation/FRAP of GFP-NRAS, palmitoylation assays, and PDE6δ co-immunoprecipitation\",\n      \"pmids\": [\"33579760\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether NRAS-specific dependence translates to differential therapeutic vulnerability in NRAS-mutant cancers was not tested\", \"Quantitative contribution of each step (palmitoylation, PDE6δ binding, Golgi flux) not resolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Establishing ICMT as a therapeutic target in progeria: genetic Icmt deletion and pharmacologic ICMT inhibition (C75) improved survival in HGPS mice and rescued senescence in HGPS cells, with on-target specificity confirmed in double-knockout controls.\",\n      \"evidence\": \"Icmt knockout in HGPS mouse model, C75 inhibitor treatment, proliferation/senescence assays, Icmt/Zmpste24 double-KO specificity control\",\n      \"pmids\": [\"33526168\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether ICMT inhibition reverses established disease or only prevents progression was not determined\", \"Potential toxicity from global ICMT inhibition in vivo not thoroughly assessed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A substrate-bound structure of eukaryotic ICMT with a prenylated peptide or full-length protein substrate has not been determined, leaving the precise geometry of methyl transfer and the structural basis for substrate selectivity among diverse prenylated proteins unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No substrate-bound eukaryotic ICMT structure\", \"Structural basis for selectivity among CAAX vs CXC substrates unknown\", \"Allosteric regulation and role of interactors such as PFKFB4 structurally uncharacterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 1, 2, 3, 5, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [1, 5, 6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [2, 4, 5, 6]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [4, 9, 11]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"KRAS\", \"NRAS\", \"RHOA\", \"RAC1\", \"PDE6D\", \"PFKFB4\"],\n    \"other_free_text\": []\n  }\n}\n```"}