{"gene":"TG_277080","gene_symbol":"MIC5","run_date":"2026-09-08T14:08:30","timeline":{"discoveries":[{"year":2000,"finding":"TgMIC5 is a member of the parvulin family of peptidyl-prolyl cis-trans isomerases (PPIases), expressed as a preproprotein processed first by signal peptidase to a proprotein, then further processed to a mature 22 kDa protein. It is stored in the micronemes of T. gondii tachyzoites and secreted into the surrounding medium.","method":"Molecular cloning, DNA sequence analysis, cell fractionation, secretion-modulating drugs, immunofluorescence microscopy, immunoelectron microscopy","journal":"Molecular and biochemical parasitology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (fractionation, IFA, immunoEM, biochemical secretion assays) in a single focused study establishing localization and processing","pmids":["11087916"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2008,"finding":"The MIC5 propeptide acts as a forward-targeting element: proteolytic maturation of MIC5 occurs at a site beyond the Golgi; only the processed form is secreted via micronemes. Deletion of the propeptide causes retention in the ER, whereas an arginine at P1' is required for correct cleavage (though P1' mutants are still cleaved at an alternative downstream site and traffic correctly). The MIC5 propeptide is interchangeable with the M2AP propeptide, indicating a common targeting mechanism.","method":"Deletion mutagenesis, site-directed mutagenesis, propeptide-swap experiments, pulse-chase/secretion assays, subcellular fractionation","journal":"Biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (mutagenesis, propeptide swaps, fractionation, secretion assays) in a single focused study","pmids":["17995454"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2006,"finding":"Genetic ablation of MIC5 enhances proteolytic trimming (by a hypothetical surface protease MPP2) of several micronemal proteins (and GRA1) on the parasite surface during invasion. MIC5 occupies the parasite surface during invasion despite lacking obvious membrane-association signals. Genetic complementation with MIC5 reverses the enhanced-processing phenotype, as does treatment with the MPP2 inhibitor ALLN. Together these data indicate MIC5 either directly inhibits MPP2 or occludes substrate cleavage sites on the parasite surface.","method":"Targeted gene deletion (knockout), genetic complementation, protease inhibitor treatment (ALLN), surface biotinylation/western blot analysis of processing, immunofluorescence","journal":"Eukaryotic cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean KO with defined molecular phenotype, complementation reversal, inhibitor validation across multiple substrates","pmids":["16980407"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2011,"finding":"A leucine at position +1 (relative to signal peptidase cleavage) of the proMIC5 propeptide is crucial for trafficking to micronemes. Propeptide domains are extensively interchangeable between T. gondii MICs and with Eimeria tenella MIC5 (EtMIC5), indicating a conserved aliphatic-residue-dependent mechanism of microneme targeting.","method":"N-terminal deletion and point mutagenesis of propeptides, propeptide-swap experiments, immunofluorescence-based trafficking assay","journal":"Traffic (Copenhagen, Denmark)","confidence":"High","confidence_rationale":"Tier 2 / Moderate — mutagenesis with multiple substitution variants and cross-species propeptide swaps, convergent results across multiple MICs","pmids":["21438967"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2012,"finding":"The 3D NMR structure of TgMIC5 reveals it structurally mimics a subtilisin prodomain, including a flexible C-terminal peptide that may insert into the active site of TgSUB1. TgMIC5 is an ~50-fold more potent inhibitor of TgSUB1 proteolytic activity than the small-molecule inhibitor ALLN. TgMIC5 is retained on the parasite plasma membrane through a direct physical interaction with the membrane-anchored TgSUB1.","method":"NMR structure determination, in vitro TgSUB1 protease activity assay with quantified IC50, co-immunoprecipitation/physical interaction assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — NMR structure plus in vitro enzymatic assay plus binding interaction, multiple orthogonal methods in a single rigorous study","pmids":["22896704"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"current_model":"TgMIC5 is a parvulin-family PPIase-like protein that is trafficked as a preproprotein to the micronemes via an aliphatic residue (Leu+1) in its propeptide — with maturation cleavage occurring post-Golgi — and, once secreted onto the parasite surface, acts as a potent (~50-fold over ALLN) inhibitor of the subtilisin protease TgSUB1 by mimicking a subtilisin prodomain, thereby suppressing trimming proteolysis of micronemal invasion proteins during host-cell entry."},"narrative":{"mechanistic_narrative":"TgMIC5 is a parvulin-family peptidyl-prolyl isomerase-like protein of Toxoplasma gondii micronemes that regulates the surface proteolysis of micronemal invasion proteins during host-cell entry [PMID:11087916, PMID:16980407]. It is synthesized as a preproprotein cleaved by signal peptidase and then matured post-Golgi to a ~22 kDa form that is stored in the micronemes and secreted [PMID:11087916]. The propeptide functions as a forward-targeting element required for exit from the ER and correct micronemal delivery, acting through an aliphatic residue (Leu at position +1) and functionally interchangeable with propeptides of other MICs and of Eimeria tenella MIC5, indicating a conserved aliphatic-residue-dependent targeting mechanism [PMID:17995454, PMID:21438967]. Once secreted onto the parasite surface, MIC5 restrains proteolytic trimming of micronemal proteins (and GRA1): its genetic ablation enhances processing, and complementation or the inhibitor ALLN reverses this phenotype [PMID:16980407]. The structural and mechanistic basis is that TgMIC5 mimics a subtilisin prodomain — its NMR structure includes a flexible C-terminal peptide poised to insert into the active site of the subtilisin protease TgSUB1 — acting as a potent inhibitor (~50-fold over ALLN) and being retained at the plasma membrane through direct physical interaction with membrane-anchored TgSUB1 [PMID:22896704].","teleology":[{"year":2000,"claim":"Establishing that MIC5 is a parvulin-family PPIase-like protein processed from a preproprotein and stored in micronemes defined it as a secreted micronemal protein of the invasion machinery.","evidence":"Molecular cloning, cell fractionation, secretion assays, immunofluorescence and immunoelectron microscopy in T. gondii tachyzoites","pmids":["11087916"],"confidence":"High","gaps":["Molecular function of the protein on the surface not yet defined","Substrate or binding partner unknown at this stage"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2006,"claim":"Genetic ablation showed MIC5 suppresses surface proteolytic trimming of micronemal proteins, revealing a regulatory role over an invasion-associated protease rather than an adhesive one.","evidence":"Targeted gene knockout, complementation, ALLN inhibitor treatment, and surface biotinylation/western analysis of processing in T. gondii","pmids":["16980407"],"confidence":"High","gaps":["Could not distinguish direct protease inhibition from substrate occlusion","Identity of the responsible protease (then termed MPP2) unresolved","Mechanism of surface retention despite lacking membrane signals unexplained"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2008,"claim":"Deletion and swap experiments demonstrated the propeptide is a forward-targeting element required for ER exit and micronemal delivery, explaining how proMIC5 is correctly routed.","evidence":"Deletion and site-directed mutagenesis, propeptide-swap with M2AP, pulse-chase/secretion assays and fractionation in T. gondii","pmids":["17995454"],"confidence":"High","gaps":["Precise trafficking receptor recognizing the propeptide not identified","Post-Golgi maturation protease not defined"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2011,"claim":"Point mutagenesis pinpointed a Leu at position +1 as the key determinant of microneme targeting and showed the mechanism is conserved across MICs and across species, refining the targeting model.","evidence":"N-terminal deletion and point mutagenesis, cross-species propeptide swaps with EtMIC5, immunofluorescence trafficking assays in T. gondii","pmids":["21438967"],"confidence":"High","gaps":["How the aliphatic residue is read out mechanistically remains unknown","The sorting machinery that recognizes it is uncharacterized"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2012,"claim":"The NMR structure and enzymatic assays resolved the mechanism: MIC5 mimics a subtilisin prodomain to directly and potently inhibit TgSUB1 and is retained on the surface via binding to it, unifying the earlier trimming and surface-retention observations.","evidence":"NMR structure determination, in vitro TgSUB1 protease activity assay with IC50 quantification, and co-immunoprecipitation in T. gondii","pmids":["22896704"],"confidence":"High","gaps":["Whether the C-terminal peptide inserts into the TgSUB1 active site not directly visualized","In vivo consequences of the MIC5–TgSUB1 interaction for invasion efficiency not quantified","Relationship of PPIase-like fold to any isomerase activity unresolved"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":null,"claim":"Whether TgMIC5 possesses genuine peptidyl-prolyl isomerase enzymatic activity, and how its protease-inhibitory role integrates quantitatively into host-cell invasion, remain open.","evidence":"","pmids":[],"confidence":"High","gaps":["No demonstration of PPIase catalytic activity in the timeline","Contribution to invasion efficiency in vivo not measured"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["TgMIC5–TgSUB1 inhibitory complex"],"partners":["TGSUB1 (SUBTILISIN PROTEASE)"],"other_free_text":["Synthesized as a preproprotein: signal peptidase cleavage to proprotein, then post-Golgi maturation to a ~22 kDa mature form [#0].","Propeptide functions as a forward-targeting element; ER exit and micronemal delivery depend on a Leu at position +1 and the propeptide is interchangeable among MICs and with Eimeria tenella MIC5 [#1, #3].","Structurally mimics a subtilisin prodomain with a flexible C-terminal peptide, inhibiting TgSUB1 ~50-fold more potently than ALLN [#4]."]},"strain_conflicts":[]},"prefetch_data":{},"corpus_meta":[{"pmid":"11087916","id":"PMC_11087916","title":"Molecular characterization of TgMIC5, a proteolytically processed antigen secreted from the micronemes of Toxoplasma gondii.","date":"2000","source":"Molecular and biochemical parasitology","url":"https://pubmed.ncbi.nlm.nih.gov/11087916","citation_count":59,"is_preprint":false},{"pmid":"12633654","id":"PMC_12633654","title":"Molecular dissection of the human B-cell response against Toxoplasma gondii infection by lambda display of cDNA libraries.","date":"2003","source":"International journal for parasitology","url":"https://pubmed.ncbi.nlm.nih.gov/12633654","citation_count":54,"is_preprint":false},{"pmid":"21438967","id":"PMC_21438967","title":"Forward targeting of Toxoplasma gondii proproteins to the micronemes involves conserved aliphatic amino acids.","date":"2011","source":"Traffic (Copenhagen, Denmark)","url":"https://pubmed.ncbi.nlm.nih.gov/21438967","citation_count":29,"is_preprint":false},{"pmid":"17995454","id":"PMC_17995454","title":"A transient forward-targeting element for microneme-regulated secretion in Toxoplasma gondii.","date":"2008","source":"Biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/17995454","citation_count":28,"is_preprint":false},{"pmid":"16980407","id":"PMC_16980407","title":"Targeted deletion of MIC5 enhances trimming proteolysis of Toxoplasma invasion proteins.","date":"2006","source":"Eukaryotic cell","url":"https://pubmed.ncbi.nlm.nih.gov/16980407","citation_count":22,"is_preprint":false},{"pmid":"22896704","id":"PMC_22896704","title":"Microneme protein 5 regulates the activity of Toxoplasma subtilisin 1 by mimicking a subtilisin prodomain.","date":"2012","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22896704","citation_count":18,"is_preprint":false},{"pmid":"34595126","id":"PMC_34595126","title":"Protective Immunity Induced by TgMIC5 and TgMIC16 DNA Vaccines Against Toxoplasmosis.","date":"2021","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/34595126","citation_count":15,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":5620,"output_tokens":1620,"usd":0.02058,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6801,"output_tokens":2871,"usd":0.05289,"stage2_stop_reason":"end_turn"},"total_usd":0.07347,"stage1_batch_id":"msgbatch_015xmQKDRqnTh9anE4m7LiD6","stage2_batch_id":"msgbatch_01JLpkFTVhoQBNgqHCKFrVVU","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2000,\n      \"finding\": \"TgMIC5 is a member of the parvulin family of peptidyl-prolyl cis-trans isomerases (PPIases), expressed as a preproprotein processed first by signal peptidase to a proprotein, then further processed to a mature 22 kDa protein. It is stored in the micronemes of T. gondii tachyzoites and secreted into the surrounding medium.\",\n      \"method\": \"Molecular cloning, DNA sequence analysis, cell fractionation, secretion-modulating drugs, immunofluorescence microscopy, immunoelectron microscopy\",\n      \"journal\": \"Molecular and biochemical parasitology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (fractionation, IFA, immunoEM, biochemical secretion assays) in a single focused study establishing localization and processing\",\n      \"pmids\": [\"11087916\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"The MIC5 propeptide acts as a forward-targeting element: proteolytic maturation of MIC5 occurs at a site beyond the Golgi; only the processed form is secreted via micronemes. Deletion of the propeptide causes retention in the ER, whereas an arginine at P1' is required for correct cleavage (though P1' mutants are still cleaved at an alternative downstream site and traffic correctly). The MIC5 propeptide is interchangeable with the M2AP propeptide, indicating a common targeting mechanism.\",\n      \"method\": \"Deletion mutagenesis, site-directed mutagenesis, propeptide-swap experiments, pulse-chase/secretion assays, subcellular fractionation\",\n      \"journal\": \"Biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (mutagenesis, propeptide swaps, fractionation, secretion assays) in a single focused study\",\n      \"pmids\": [\"17995454\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Genetic ablation of MIC5 enhances proteolytic trimming (by a hypothetical surface protease MPP2) of several micronemal proteins (and GRA1) on the parasite surface during invasion. MIC5 occupies the parasite surface during invasion despite lacking obvious membrane-association signals. Genetic complementation with MIC5 reverses the enhanced-processing phenotype, as does treatment with the MPP2 inhibitor ALLN. Together these data indicate MIC5 either directly inhibits MPP2 or occludes substrate cleavage sites on the parasite surface.\",\n      \"method\": \"Targeted gene deletion (knockout), genetic complementation, protease inhibitor treatment (ALLN), surface biotinylation/western blot analysis of processing, immunofluorescence\",\n      \"journal\": \"Eukaryotic cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO with defined molecular phenotype, complementation reversal, inhibitor validation across multiple substrates\",\n      \"pmids\": [\"16980407\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"A leucine at position +1 (relative to signal peptidase cleavage) of the proMIC5 propeptide is crucial for trafficking to micronemes. Propeptide domains are extensively interchangeable between T. gondii MICs and with Eimeria tenella MIC5 (EtMIC5), indicating a conserved aliphatic-residue-dependent mechanism of microneme targeting.\",\n      \"method\": \"N-terminal deletion and point mutagenesis of propeptides, propeptide-swap experiments, immunofluorescence-based trafficking assay\",\n      \"journal\": \"Traffic (Copenhagen, Denmark)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mutagenesis with multiple substitution variants and cross-species propeptide swaps, convergent results across multiple MICs\",\n      \"pmids\": [\"21438967\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"The 3D NMR structure of TgMIC5 reveals it structurally mimics a subtilisin prodomain, including a flexible C-terminal peptide that may insert into the active site of TgSUB1. TgMIC5 is an ~50-fold more potent inhibitor of TgSUB1 proteolytic activity than the small-molecule inhibitor ALLN. TgMIC5 is retained on the parasite plasma membrane through a direct physical interaction with the membrane-anchored TgSUB1.\",\n      \"method\": \"NMR structure determination, in vitro TgSUB1 protease activity assay with quantified IC50, co-immunoprecipitation/physical interaction assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — NMR structure plus in vitro enzymatic assay plus binding interaction, multiple orthogonal methods in a single rigorous study\",\n      \"pmids\": [\"22896704\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"current_model\": \"TgMIC5 is a parvulin-family PPIase-like protein that is trafficked as a preproprotein to the micronemes via an aliphatic residue (Leu+1) in its propeptide — with maturation cleavage occurring post-Golgi — and, once secreted onto the parasite surface, acts as a potent (~50-fold over ALLN) inhibitor of the subtilisin protease TgSUB1 by mimicking a subtilisin prodomain, thereby suppressing trimming proteolysis of micronemal invasion proteins during host-cell entry.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"TgMIC5 is a parvulin-family peptidyl-prolyl isomerase-like protein of Toxoplasma gondii micronemes that regulates the surface proteolysis of micronemal invasion proteins during host-cell entry [#0, #2]. It is synthesized as a preproprotein cleaved by signal peptidase and then matured post-Golgi to a ~22 kDa form that is stored in the micronemes and secreted [#0]. The propeptide functions as a forward-targeting element required for exit from the ER and correct micronemal delivery, acting through an aliphatic residue (Leu at position +1) and functionally interchangeable with propeptides of other MICs and of Eimeria tenella MIC5, indicating a conserved aliphatic-residue-dependent targeting mechanism [#1, #3]. Once secreted onto the parasite surface, MIC5 restrains proteolytic trimming of micronemal proteins (and GRA1): its genetic ablation enhances processing, and complementation or the inhibitor ALLN reverses this phenotype [#2]. The structural and mechanistic basis is that TgMIC5 mimics a subtilisin prodomain \\u2014 its NMR structure includes a flexible C-terminal peptide poised to insert into the active site of the subtilisin protease TgSUB1 \\u2014 acting as a potent inhibitor (~50-fold over ALLN) and being retained at the plasma membrane through direct physical interaction with membrane-anchored TgSUB1 [#4].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Establishing that MIC5 is a parvulin-family PPIase-like protein processed from a preproprotein and stored in micronemes defined it as a secreted micronemal protein of the invasion machinery.\",\n      \"evidence\": \"Molecular cloning, cell fractionation, secretion assays, immunofluorescence and immunoelectron microscopy in T. gondii tachyzoites\",\n      \"pmids\": [\"11087916\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular function of the protein on the surface not yet defined\", \"Substrate or binding partner unknown at this stage\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Genetic ablation showed MIC5 suppresses surface proteolytic trimming of micronemal proteins, revealing a regulatory role over an invasion-associated protease rather than an adhesive one.\",\n      \"evidence\": \"Targeted gene knockout, complementation, ALLN inhibitor treatment, and surface biotinylation/western analysis of processing in T. gondii\",\n      \"pmids\": [\"16980407\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Could not distinguish direct protease inhibition from substrate occlusion\", \"Identity of the responsible protease (then termed MPP2) unresolved\", \"Mechanism of surface retention despite lacking membrane signals unexplained\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Deletion and swap experiments demonstrated the propeptide is a forward-targeting element required for ER exit and micronemal delivery, explaining how proMIC5 is correctly routed.\",\n      \"evidence\": \"Deletion and site-directed mutagenesis, propeptide-swap with M2AP, pulse-chase/secretion assays and fractionation in T. gondii\",\n      \"pmids\": [\"17995454\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Precise trafficking receptor recognizing the propeptide not identified\", \"Post-Golgi maturation protease not defined\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Point mutagenesis pinpointed a Leu at position +1 as the key determinant of microneme targeting and showed the mechanism is conserved across MICs and across species, refining the targeting model.\",\n      \"evidence\": \"N-terminal deletion and point mutagenesis, cross-species propeptide swaps with EtMIC5, immunofluorescence trafficking assays in T. gondii\",\n      \"pmids\": [\"21438967\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How the aliphatic residue is read out mechanistically remains unknown\", \"The sorting machinery that recognizes it is uncharacterized\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"The NMR structure and enzymatic assays resolved the mechanism: MIC5 mimics a subtilisin prodomain to directly and potently inhibit TgSUB1 and is retained on the surface via binding to it, unifying the earlier trimming and surface-retention observations.\",\n      \"evidence\": \"NMR structure determination, in vitro TgSUB1 protease activity assay with IC50 quantification, and co-immunoprecipitation in T. gondii\",\n      \"pmids\": [\"22896704\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether the C-terminal peptide inserts into the TgSUB1 active site not directly visualized\", \"In vivo consequences of the MIC5\\u2013TgSUB1 interaction for invasion efficiency not quantified\", \"Relationship of PPIase-like fold to any isomerase activity unresolved\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Whether TgMIC5 possesses genuine peptidyl-prolyl isomerase enzymatic activity, and how its protease-inhibitory role integrates quantitatively into host-cell invasion, remain open.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No demonstration of PPIase catalytic activity in the timeline\", \"Contribution to invasion efficiency in vivo not measured\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0004866\", \"supporting_discovery_ids\": [2, 4], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0003755\", \"supporting_discovery_ids\": [0], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0020009\", \"supporting_discovery_ids\": [0], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0009986\", \"supporting_discovery_ids\": [2, 4], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [0], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0044409\", \"supporting_discovery_ids\": [2, 4], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"complexes\": [\"TgMIC5\\u2013TgSUB1 inhibitory complex\"],\n    \"partners\": [\"TgSUB1 (subtilisin protease)\"],\n    \"other_free_text\": [\n      \"Synthesized as a preproprotein: signal peptidase cleavage to proprotein, then post-Golgi maturation to a ~22 kDa mature form [#0].\",\n      \"Propeptide functions as a forward-targeting element; ER exit and micronemal delivery depend on a Leu at position +1 and the propeptide is interchangeable among MICs and with Eimeria tenella MIC5 [#1, #3].\",\n      \"Structurally mimics a subtilisin prodomain with a flexible C-terminal peptide, inhibiting TgSUB1 ~50-fold more potently than ALLN [#4].\"\n    ]\n  },\n  \"strain_conflicts\": []\n}\n```","audit_flag":null,"strains":[{"strain":"ME49","gene_id":"TGME49_277080","name":"MIC5","product":"microneme protein MIC5","uniprot_acc":"S8GAQ8","chromosome":"TGME49_chrXII","start_pos":6723174},{"strain":"GT1","gene_id":"TGGT1_277080","name":null,"product":"microneme protein MIC5","uniprot_acc":null,"chromosome":"TGGT1_chrXII","start_pos":6631512},{"strain":"RH88","gene_id":"TGRH88_066240","name":null,"product":"microneme protein MIC5","uniprot_acc":null,"chromosome":"CM023093","start_pos":6804523},{"strain":"VEG","gene_id":"TGVEG_277080","name":null,"product":"microneme protein MIC5","uniprot_acc":"A0A125YHB4","chromosome":"TGVEG_chrXII","start_pos":6740775}]}