{"gene":"PF3D7_1232100","run_date":"2026-09-08T14:19:06","timeline":{"discoveries":[{"year":1994,"finding":"CPN60 (PF3D7_1232100) was isolated and sequenced; the gene is located on chromosome 12, contains a single intron of 868 nucleotides (the largest then identified in P. falciparum), encodes a predicted 719 amino acid / 81.6 kDa protein with a putative mitochondrial signal peptide and an unusually long carboxy-terminal extension composed almost entirely of glutamic and aspartic acid residues, and produces a 4-kb transcript.","method":"Genomic cloning, sequencing, Southern and Northern blotting","journal":"Molecular and biochemical parasitology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct genomic/sequence characterisation with multiple methods (cloning, sequencing, blotting) in a single study; no functional reconstitution","pmids":["7915821"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2005,"finding":"P. falciparum possesses two Cpn60 proteins, one targeted exclusively to the mitochondrion and the other exclusively to the apicoplast (plastid), each forming a distinct type I chaperonin system with its own organelle-specific cochaperonin (PfCpn10 in mitochondrion, PfCpn20 in plastid). Transfection experiments with fluorescent reporters established the organelle-specific localisations.","method":"Transfection with fluorescent reporter constructs (targeting-sequence fusions), comparative sequence and phylogenetic analysis","journal":"Molecular and biochemical parasitology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct subcellular localisation by live fluorescence imaging with targeting constructs, single lab, complemented by phylogenetic analysis","pmids":["15926202"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2021,"finding":"Crystal structure of P. falciparum mitochondrial CPN60 (PF3D7_1232100) bound to ATP was determined, revealing a double heptameric ring assembly in a unique open/dynamic intermediate state not seen in other Cpn60 structures. Three of the seven apical domains are highly dynamic while the equatorial domains form a stable ring. A unique 26–27 residue insertion in the equatorial domain greatly increases both inter-ring and intra-ring subunit–subunit interactions. The structure implies large apical-domain movements driven by nucleotide binding regulate substrate-binding and folding.","method":"X-ray crystallography (crystal structure of ATP-bound complex), structural comparative analysis","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 1 / Moderate — atomic-resolution crystal structure with bound nucleotide; single lab but direct structural evidence with mechanistic interpretation of conformational dynamics and domain interactions","pmids":["33723304"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2025,"finding":"Genetic ablation of the apicoplast CPN60 subunit (PF3D7_1232100) in P. falciparum causes parasite death from organellar damage within the first replication cycle — distinct from the delayed-death phenotype of apicoplast translation inhibitors — establishing an essential role in organelle biogenesis and proteostasis. Unlike prokaryotic GroEL orthologues, CPN60 is not upregulated during heat shock and does not affect the heat-shock response.","method":"Genetic ablation (conditional knockout), parasite viability assays, heat-shock assays","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean loss-of-function with defined cellular phenotype and specific control experiments (heat-shock), single lab","pmids":["40694565"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2025,"finding":"CPN60 (PF3D7_1232100) physically binds both the active and inactive forms of the Clp (caseinolytic protease) proteolytic complex in the apicoplast, manipulating its stability; a computational structural model supports a stable interface between the two large complexes. This interaction links chaperonin-mediated refolding to proteolytic control of organelle proteostasis.","method":"Co-immunoprecipitation / physical interaction assays, computational structural modelling","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — direct binding shown by co-IP of both active and inactive Clp forms, supported by structural modelling, single lab; no in vitro reconstitution","pmids":["40694565"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"current_model":"P. falciparum CPN60 (PF3D7_1232100) is an essential apicoplast-localised type I chaperonin that assembles as an ATP-bound double heptameric ring with a unique equatorial insertion enhancing inter-subunit contacts and dynamic apical domains for substrate binding; it is required for apicoplast integrity and parasite survival within a single replication cycle, and unlike its bacterial orthologue GroEL it is not heat-shock regulated but instead directly binds and stabilises the Clp proteolytic complex, coupling protein refolding to proteolysis to safeguard organelle proteostasis."},"narrative":{"mechanistic_narrative":"CPN60 (PF3D7_1232100) is an essential P. falciparum type I chaperonin required for organelle proteostasis and parasite survival [PMID:40694565]. The gene, located on chromosome 12 with a single unusually long intron, encodes a ~81.6 kDa protein bearing an organelle-targeting signal peptide and a distinctive acidic (glutamate/aspartate-rich) carboxy-terminal extension [PMID:7915821]. P. falciparum encodes two Cpn60 proteins that partition between the mitochondrion and the apicoplast, each pairing with its own organelle-specific cochaperonin [PMID:15926202]. Structurally, ATP-bound CPN60 assembles into a double heptameric ring captured in an open, dynamic intermediate state in which three apical domains are mobile while the equatorial domains form a stable ring; a unique 26–27 residue equatorial insertion strengthens both inter- and intra-ring subunit contacts, and nucleotide-driven apical-domain movements are inferred to govern substrate binding and folding [PMID:33723304]. Genetic ablation of the apicoplast CPN60 subunit kills parasites within the first replication cycle through organellar damage, a rapid-death phenotype distinct from the delayed death of apicoplast translation inhibitors, and unlike bacterial GroEL orthologues CPN60 is not upregulated by heat shock [PMID:40694565]. CPN60 physically binds both active and inactive forms of the apicoplast Clp proteolytic complex, coupling chaperonin-mediated refolding to proteolytic control of organelle proteostasis [PMID:40694565].","teleology":[{"year":1994,"claim":"Establishing the existence and primary structure of the gene was the first step, defining CPN60 as a chaperonin-family protein with unusual features (a large intron, an acidic C-terminal tail, and an organelle-targeting signal) that distinguished it from typical orthologues.","evidence":"Genomic cloning, sequencing, and Southern/Northern blotting in P. falciparum","pmids":["7915821"],"confidence":"Medium","gaps":["Subcellular localisation was only predicted from a putative signal peptide, not shown experimentally","No functional or biochemical activity was demonstrated","The role of the acidic C-terminal extension was not addressed"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2005,"claim":"Resolving where CPN60 acts, this work showed P. falciparum carries two distinct Cpn60 systems partitioned between mitochondrion and apicoplast, each with a dedicated cochaperonin, clarifying that chaperonin function is compartmentalised across the two endosymbiotic organelles.","evidence":"Transfection with fluorescent targeting-sequence reporters plus phylogenetic analysis in P. falciparum","pmids":["15926202"],"confidence":"Medium","gaps":["Reporter localisation with targeting sequences does not confirm the native full-length protein's destination","No demonstration of chaperonin folding activity in either compartment","Functional distinction between the two Cpn60 systems not tested"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2021,"claim":"The ATP-bound crystal structure answered how CPN60 is assembled and how it might work mechanically, revealing a double heptameric ring in a dynamic intermediate state and a unique equatorial insertion that reinforces subunit contacts, linking nucleotide binding to apical-domain motions that govern substrate handling.","evidence":"X-ray crystallography of the ATP-bound complex with structural comparative analysis","pmids":["33723304"],"confidence":"High","gaps":["No substrate or folding cycle captured; substrate-binding inferred from domain dynamics","Functional consequence of the equatorial insertion not tested by mutagenesis","Structure does not establish the cochaperonin-bound state"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2025,"claim":"Conditional ablation established CPN60 as essential for apicoplast biogenesis and proteostasis, and the rapid single-cycle death distinct from delayed-death translation inhibitors, together with the lack of heat-shock upregulation, showed its role diverges from bacterial GroEL.","evidence":"Conditional knockout, parasite viability assays, and heat-shock assays in P. falciparum","pmids":["40694565"],"confidence":"Medium","gaps":["The specific client proteins whose misfolding causes death were not identified","Mechanism linking organellar damage to rapid death not resolved","Single-lab loss-of-function without complementation rescue reported here"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2025,"claim":"Identifying a physical interaction with the Clp proteolytic complex provided a mechanistic link between chaperonin refolding and proteolysis, indicating CPN60 helps coordinate the balance between protein folding and degradation to safeguard apicoplast proteostasis.","evidence":"Co-immunoprecipitation of active and inactive Clp forms plus computational structural modelling in P. falciparum","pmids":["40694565"],"confidence":"Medium","gaps":["Interaction shown by co-IP and modelling without in vitro reconstitution","Functional outcome of the CPN60–Clp interaction on substrate fate not demonstrated","Interface predicted computationally, not structurally resolved"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":null,"claim":"The physiological substrates folded by CPN60 and the biochemical mechanism by which it stabilises versus activates the Clp complex remain undefined.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No identified endogenous folding clients","No in vitro reconstitution of the chaperonin folding cycle or of CPN60–Clp coupling","Functional role of the acidic C-terminal extension uncharacterised"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["Double heptameric CPN60 chaperonin ring (ATP-bound) [#2]","CPN60–Clp proteolytic complex assembly [#4]"],"partners":["CLP CASEINOLYTIC PROTEASE COMPLEX (ACTIVE AND INACTIVE FORMS) [#4]","ORGANELLE-SPECIFIC COCHAPERONINS PFCPN10 (MITOCHONDRION) AND PFCPN20 (APICOPLAST) [#1]"],"other_free_text":["Encoded on chromosome 12 with a single ~868 nt intron; predicted ~81.6 kDa / 719 aa protein with an organelle-targeting signal peptide and an acidic (Glu/Asp-rich) C-terminal extension [#0]","Contains a unique 26–27 residue equatorial-domain insertion that strengthens inter- and intra-ring subunit contacts [#2]","Not heat-shock inducible, unlike bacterial GroEL orthologues [#3]","Ablation causes rapid single-cycle parasite death, distinct from apicoplast delayed-death phenotype [#3]"]}},"prefetch_data":{},"corpus_meta":[{"pmid":"14747157","id":"PMC_14747157","title":"The use of DsRED in single- and dual-color fluorescence labeling of mitochondrial and plastid organelles in Plasmodium falciparum.","date":"2004","source":"Molecular and biochemical parasitology","url":"https://pubmed.ncbi.nlm.nih.gov/14747157","citation_count":25,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["curated_ref"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"7915821","id":"PMC_7915821","title":"Isolation and characterization of a chaperonin-60 gene of the human malaria parasite Plasmodium falciparum.","date":"1994","source":"Molecular and biochemical parasitology","url":"https://pubmed.ncbi.nlm.nih.gov/7915821","citation_count":23,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["curated_ref"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"15926202","id":"PMC_15926202","title":"Organelle-specific cochaperonins in apicomplexan parasites.","date":"2005","source":"Molecular and biochemical parasitology","url":"https://pubmed.ncbi.nlm.nih.gov/15926202","citation_count":22,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["CPN60"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"33723304","id":"PMC_33723304","title":"Crystal structure of P. falciparum Cpn60 bound to ATP reveals an open dynamic conformation before substrate binding.","date":"2021","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/33723304","citation_count":4,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["CPN60"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"40694565","id":"PMC_40694565","title":"A chaperonin complex regulates organelle proteostasis in malaria parasites.","date":"2025","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/40694565","citation_count":2,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["CPN60"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":4612,"output_tokens":1546,"usd":0.018513,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6617,"output_tokens":3030,"usd":0.054418,"stage2_stop_reason":"end_turn"},"total_usd":0.072931,"stage1_batch_id":"msgbatch_011BPjN2BL33F8cJXD1r7V7T","stage2_batch_id":"msgbatch_01Qwg6vP7qxkpL6Gsc7tajsb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1994,\n      \"finding\": \"CPN60 (PF3D7_1232100) was isolated and sequenced; the gene is located on chromosome 12, contains a single intron of 868 nucleotides (the largest then identified in P. falciparum), encodes a predicted 719 amino acid / 81.6 kDa protein with a putative mitochondrial signal peptide and an unusually long carboxy-terminal extension composed almost entirely of glutamic and aspartic acid residues, and produces a 4-kb transcript.\",\n      \"method\": \"Genomic cloning, sequencing, Southern and Northern blotting\",\n      \"journal\": \"Molecular and biochemical parasitology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct genomic/sequence characterisation with multiple methods (cloning, sequencing, blotting) in a single study; no functional reconstitution\",\n      \"pmids\": [\"7915821\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"P. falciparum possesses two Cpn60 proteins, one targeted exclusively to the mitochondrion and the other exclusively to the apicoplast (plastid), each forming a distinct type I chaperonin system with its own organelle-specific cochaperonin (PfCpn10 in mitochondrion, PfCpn20 in plastid). Transfection experiments with fluorescent reporters established the organelle-specific localisations.\",\n      \"method\": \"Transfection with fluorescent reporter constructs (targeting-sequence fusions), comparative sequence and phylogenetic analysis\",\n      \"journal\": \"Molecular and biochemical parasitology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct subcellular localisation by live fluorescence imaging with targeting constructs, single lab, complemented by phylogenetic analysis\",\n      \"pmids\": [\"15926202\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Crystal structure of P. falciparum mitochondrial CPN60 (PF3D7_1232100) bound to ATP was determined, revealing a double heptameric ring assembly in a unique open/dynamic intermediate state not seen in other Cpn60 structures. Three of the seven apical domains are highly dynamic while the equatorial domains form a stable ring. A unique 26–27 residue insertion in the equatorial domain greatly increases both inter-ring and intra-ring subunit–subunit interactions. The structure implies large apical-domain movements driven by nucleotide binding regulate substrate-binding and folding.\",\n      \"method\": \"X-ray crystallography (crystal structure of ATP-bound complex), structural comparative analysis\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — atomic-resolution crystal structure with bound nucleotide; single lab but direct structural evidence with mechanistic interpretation of conformational dynamics and domain interactions\",\n      \"pmids\": [\"33723304\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Genetic ablation of the apicoplast CPN60 subunit (PF3D7_1232100) in P. falciparum causes parasite death from organellar damage within the first replication cycle — distinct from the delayed-death phenotype of apicoplast translation inhibitors — establishing an essential role in organelle biogenesis and proteostasis. Unlike prokaryotic GroEL orthologues, CPN60 is not upregulated during heat shock and does not affect the heat-shock response.\",\n      \"method\": \"Genetic ablation (conditional knockout), parasite viability assays, heat-shock assays\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean loss-of-function with defined cellular phenotype and specific control experiments (heat-shock), single lab\",\n      \"pmids\": [\"40694565\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"CPN60 (PF3D7_1232100) physically binds both the active and inactive forms of the Clp (caseinolytic protease) proteolytic complex in the apicoplast, manipulating its stability; a computational structural model supports a stable interface between the two large complexes. This interaction links chaperonin-mediated refolding to proteolytic control of organelle proteostasis.\",\n      \"method\": \"Co-immunoprecipitation / physical interaction assays, computational structural modelling\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — direct binding shown by co-IP of both active and inactive Clp forms, supported by structural modelling, single lab; no in vitro reconstitution\",\n      \"pmids\": [\"40694565\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    }\n  ],\n  \"current_model\": \"P. falciparum CPN60 (PF3D7_1232100) is an essential apicoplast-localised type I chaperonin that assembles as an ATP-bound double heptameric ring with a unique equatorial insertion enhancing inter-subunit contacts and dynamic apical domains for substrate binding; it is required for apicoplast integrity and parasite survival within a single replication cycle, and unlike its bacterial orthologue GroEL it is not heat-shock regulated but instead directly binds and stabilises the Clp proteolytic complex, coupling protein refolding to proteolysis to safeguard organelle proteostasis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CPN60 (PF3D7_1232100) is an essential P. falciparum type I chaperonin required for organelle proteostasis and parasite survival [#3]. The gene, located on chromosome 12 with a single unusually long intron, encodes a ~81.6 kDa protein bearing an organelle-targeting signal peptide and a distinctive acidic (glutamate/aspartate-rich) carboxy-terminal extension [#0]. P. falciparum encodes two Cpn60 proteins that partition between the mitochondrion and the apicoplast, each pairing with its own organelle-specific cochaperonin [#1]. Structurally, ATP-bound CPN60 assembles into a double heptameric ring captured in an open, dynamic intermediate state in which three apical domains are mobile while the equatorial domains form a stable ring; a unique 26–27 residue equatorial insertion strengthens both inter- and intra-ring subunit contacts, and nucleotide-driven apical-domain movements are inferred to govern substrate binding and folding [#2]. Genetic ablation of the apicoplast CPN60 subunit kills parasites within the first replication cycle through organellar damage, a rapid-death phenotype distinct from the delayed death of apicoplast translation inhibitors, and unlike bacterial GroEL orthologues CPN60 is not upregulated by heat shock [#3]. CPN60 physically binds both active and inactive forms of the apicoplast Clp proteolytic complex, coupling chaperonin-mediated refolding to proteolytic control of organelle proteostasis [#4].\",\n  \"teleology\": [\n    {\n      \"year\": 1994,\n      \"claim\": \"Establishing the existence and primary structure of the gene was the first step, defining CPN60 as a chaperonin-family protein with unusual features (a large intron, an acidic C-terminal tail, and an organelle-targeting signal) that distinguished it from typical orthologues.\",\n      \"evidence\": \"Genomic cloning, sequencing, and Southern/Northern blotting in P. falciparum\",\n      \"pmids\": [\"7915821\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Subcellular localisation was only predicted from a putative signal peptide, not shown experimentally\",\n        \"No functional or biochemical activity was demonstrated\",\n        \"The role of the acidic C-terminal extension was not addressed\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Resolving where CPN60 acts, this work showed P. falciparum carries two distinct Cpn60 systems partitioned between mitochondrion and apicoplast, each with a dedicated cochaperonin, clarifying that chaperonin function is compartmentalised across the two endosymbiotic organelles.\",\n      \"evidence\": \"Transfection with fluorescent targeting-sequence reporters plus phylogenetic analysis in P. falciparum\",\n      \"pmids\": [\"15926202\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Reporter localisation with targeting sequences does not confirm the native full-length protein's destination\",\n        \"No demonstration of chaperonin folding activity in either compartment\",\n        \"Functional distinction between the two Cpn60 systems not tested\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"The ATP-bound crystal structure answered how CPN60 is assembled and how it might work mechanically, revealing a double heptameric ring in a dynamic intermediate state and a unique equatorial insertion that reinforces subunit contacts, linking nucleotide binding to apical-domain motions that govern substrate handling.\",\n      \"evidence\": \"X-ray crystallography of the ATP-bound complex with structural comparative analysis\",\n      \"pmids\": [\"33723304\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No substrate or folding cycle captured; substrate-binding inferred from domain dynamics\",\n        \"Functional consequence of the equatorial insertion not tested by mutagenesis\",\n        \"Structure does not establish the cochaperonin-bound state\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Conditional ablation established CPN60 as essential for apicoplast biogenesis and proteostasis, and the rapid single-cycle death distinct from delayed-death translation inhibitors, together with the lack of heat-shock upregulation, showed its role diverges from bacterial GroEL.\",\n      \"evidence\": \"Conditional knockout, parasite viability assays, and heat-shock assays in P. falciparum\",\n      \"pmids\": [\"40694565\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"The specific client proteins whose misfolding causes death were not identified\",\n        \"Mechanism linking organellar damage to rapid death not resolved\",\n        \"Single-lab loss-of-function without complementation rescue reported here\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identifying a physical interaction with the Clp proteolytic complex provided a mechanistic link between chaperonin refolding and proteolysis, indicating CPN60 helps coordinate the balance between protein folding and degradation to safeguard apicoplast proteostasis.\",\n      \"evidence\": \"Co-immunoprecipitation of active and inactive Clp forms plus computational structural modelling in P. falciparum\",\n      \"pmids\": [\"40694565\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Interaction shown by co-IP and modelling without in vitro reconstitution\",\n        \"Functional outcome of the CPN60–Clp interaction on substrate fate not demonstrated\",\n        \"Interface predicted computationally, not structurally resolved\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The physiological substrates folded by CPN60 and the biochemical mechanism by which it stabilises versus activates the Clp complex remain undefined.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No identified endogenous folding clients\",\n        \"No in vitro reconstitution of the chaperonin folding cycle or of CPN60–Clp coupling\",\n        \"Functional role of the acidic C-terminal extension uncharacterised\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140662\", \"supporting_discovery_ids\": [2, 3]},\n      {\"term_id\": \"GO:0005524\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0051082\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0020011\", \"supporting_discovery_ids\": [1, 3]},\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0006457\", \"supporting_discovery_ids\": [2, 3, 4]},\n      {\"term_id\": \"GO:0009658\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"complexes\": [\n      \"Double heptameric CPN60 chaperonin ring (ATP-bound) [#2]\",\n      \"CPN60–Clp proteolytic complex assembly [#4]\"\n    ],\n    \"partners\": [\n      \"Clp caseinolytic protease complex (active and inactive forms) [#4]\",\n      \"Organelle-specific cochaperonins PfCpn10 (mitochondrion) and PfCpn20 (apicoplast) [#1]\"\n    ],\n    \"other_free_text\": [\n      \"Encoded on chromosome 12 with a single ~868 nt intron; predicted ~81.6 kDa / 719 aa protein with an organelle-targeting signal peptide and an acidic (Glu/Asp-rich) C-terminal extension [#0]\",\n      \"Contains a unique 26–27 residue equatorial-domain insertion that strengthens inter- and intra-ring subunit contacts [#2]\",\n      \"Not heat-shock inducible, unlike bacterial GroEL orthologues [#3]\",\n      \"Ablation causes rapid single-cycle parasite death, distinct from apicoplast delayed-death phenotype [#3]\"\n    ]\n  }\n}","audit_flag":null,"strains":[{"strain":"Pf3D7","gene_id":"PF3D7_1232100","name":null,"product":"60 kDa chaperonin","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PbANKA","gene_id":"PBANKA_1446800","name":null,"product":"60 kDa chaperonin, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PkH","gene_id":"PKNH_1451600","name":null,"product":"60 kDa chaperonin, putative","uniprot_acc":null,"chromosome":null,"start_pos":null}],"gene_symbol":"CPN60","evaluation":null}