{"gene":"PF3D7_0528100","run_date":"2026-09-07T10:55:52","timeline":{"discoveries":[{"year":2020,"finding":"AP-2μ (the μ subunit of the AP-2 adaptin complex in P. falciparum) is essential for blood-stage replication: conditional gene knockout severely disrupted schizont organization and maturation and caused mislocalization of key merozoite proteins.","method":"Conditional gene knockout with fluorescence microscopy phenotypic readout","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 / Moderate — conditional KO with defined cellular phenotype (schizont disruption, merozoite protein mislocalization), single lab but multiple orthogonal methods (KO + fluorescence + electron microscopy)","pmids":["32098816"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2020,"finding":"HA-tagged AP-2μ localizes in P. falciparum as determined by fluorescence and electron microscopy, placing it in a cellular compartment consistent with a trafficking/vesicular function.","method":"Fluorescence microscopy and electron microscopy of HA-tagged AP-2μ transgenic parasites","journal":"mBio","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — direct localization by two imaging modalities in a single lab, but no functional consequence specifically linked to localization site","pmids":["32098816"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2020,"finding":"Co-immunoprecipitation and mass spectrometry identified AP-2μ-interacting partners including other AP-2 subunits (consistent with a heterotetrameric AP-2 complex), the K10 kelch-domain protein, and PfEHD (an effector of endocytosis and lipid mobilization).","method":"Co-immunoprecipitation of HA-tagged AP-2μ followed by mass spectrometry","journal":"mBio","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP with MS identification, single lab, multiple interactors identified","pmids":["32098816"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2020,"finding":"No interaction between AP-2 complex subunits and clathrin was detected; reverse immunoprecipitation with a clathrin nanobody pulled down other heterotetrameric AP-complexes but not AP-2 subunits, indicating the P. falciparum AP-2 complex operates independently of clathrin.","method":"Reverse immunoprecipitation using a clathrin nanobody","journal":"mBio","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — negative result established by reciprocal Co-IP with nanobody, single lab; negative finding is mechanistically informative","pmids":["32098816"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"current_model":"In P. falciparum, the AP-2 complex β subunit (PF3D7_0528100) is part of a heterotetrameric AP-2 adaptin complex that is essential for schizont maturation and merozoite protein localization during blood-stage replication; the complex interacts with other AP-2 subunits, the K10 kelch-domain protein, and PfEHD, but—unlike canonical AP-2 in other organisms—operates independently of clathrin, indicating a specialized, non-classical endocytic or trafficking function in malaria parasites."},"narrative":{"mechanistic_narrative":"PF3D7_0528100 is annotated as the putative β subunit of the P. falciparum AP-1/2 adaptin complex, but the available corpus characterizes it only through its membership in the heterotetrameric AP-2 complex rather than through experiments performed on the β subunit itself. The functional studies in this corpus were carried out on the AP-2 μ subunit, whose conditional knockout disrupts schizont organization and maturation and mislocalizes merozoite proteins, establishing the AP-2 complex as essential for blood-stage replication [PMID:32098816]. Co-immunoprecipitation and mass spectrometry of tagged AP-2μ recovered other AP-2 subunits—consistent with an assembled heterotetramer that would include the β subunit—along with the K10 kelch-domain protein and the endocytic effector PfEHD [PMID:32098816], and localization imaging places the complex in a compartment consistent with a vesicular/trafficking role [PMID:32098816]. Notably, reverse immunoprecipitation with a clathrin nanobody failed to recover AP-2 subunits while pulling down other AP complexes, indicating that the P. falciparum AP-2 complex operates independently of clathrin, unlike canonical AP-2 elsewhere [PMID:32098816]. Beyond its inferred membership in this complex, no experiment in the available corpus directly assays PF3D7_0528100 itself—its individual function, essentiality, and interactions have not been characterized.","teleology":[{"year":2020,"claim":"Whether the P. falciparum AP-2 complex is required for blood-stage growth was unknown; conditional knockout of the AP-2 μ subunit showed the complex is essential for schizont maturation and correct merozoite protein localization, placing AP-2 in the replicative program.","evidence":"Conditional gene knockout of AP-2μ with fluorescence and electron microscopy phenotyping","pmids":["32098816"],"confidence":"High","gaps":["The β subunit PF3D7_0528100 itself was not knocked out; essentiality is demonstrated only for the μ subunit","The precise trafficking cargo and mechanistic step disrupted are not defined","No individual functional assay of PF3D7_0528100"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2020,"claim":"The subcellular context of the AP-2 complex was undefined; imaging of tagged AP-2μ localized it to a compartment consistent with vesicular trafficking, supporting a membrane-associated adaptor role.","evidence":"Fluorescence and electron microscopy of HA-tagged AP-2μ transgenic parasites","pmids":["32098816"],"confidence":"Medium","gaps":["Localization was shown for the μ subunit, not for PF3D7_0528100","No functional consequence was tied to the specific localization site"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2020,"claim":"The composition and partners of the complex were unknown; Co-IP/MS of AP-2μ recovered other AP-2 subunits (implying the β subunit assembles into the heterotetramer), the K10 kelch protein, and PfEHD, defining an interaction network for the complex.","evidence":"Co-immunoprecipitation of HA-tagged AP-2μ followed by mass spectrometry","pmids":["32098816"],"confidence":"Medium","gaps":["PF3D7_0528100 is inferred as a complex member but not independently verified by reciprocal pulldown from the β subunit","Functional roles of the K10 and PfEHD interactions are not established","Single lab, single Co-IP bait"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2020,"claim":"Whether the parasite AP-2 complex uses the canonical clathrin-dependent pathway was untested; reverse Co-IP with a clathrin nanobody recovered other AP complexes but not AP-2, establishing that P. falciparum AP-2 acts independently of clathrin.","evidence":"Reverse immunoprecipitation using a clathrin nanobody","pmids":["32098816"],"confidence":"Medium","gaps":["Negative result does not identify what alternative coat or machinery AP-2 uses","Interaction tested via the μ subunit context, not PF3D7_0528100 directly"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":null,"claim":"No experiment has directly assayed PF3D7_0528100 (the β subunit) itself; its individual contribution to complex assembly, cargo selection, and the clathrin-independent trafficking mechanism remains unknown.","evidence":"","pmids":[],"confidence":"Low","gaps":["No knockout, tagging, or interaction study performed on PF3D7_0528100 specifically","Molecular function of the β subunit is inferred solely from complex membership","The nature of the clathrin-independent trafficking pathway is undefined"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["AP-2 adaptor complex (heterotetrameric)"],"partners":["AP-2 COMPLEX SUBUNITS","K10 KELCH-DOMAIN PROTEIN","PFEHD"],"other_free_text":["PF3D7_0528100 is characterized in the corpus only as an inferred β-subunit member of the P. falciparum AP-2 complex; all functional, localization, and interaction data derive from experiments on the AP-2 μ subunit, not the β subunit itself.","The P. falciparum AP-2 complex operates independently of clathrin [#3], distinguishing it from canonical AP-2 adaptors.","AP-2 complex essentiality was demonstrated for schizont maturation and merozoite protein localization during blood-stage replication [#0]."]}},"prefetch_data":{},"corpus_meta":[{"pmid":"32098816","id":"PMC_32098816","title":"The Plasmodium falciparum Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont Maturation.","date":"2020","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/32098816","citation_count":31,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["curated_ref"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":3467,"output_tokens":1048,"usd":0.01306,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":5828,"output_tokens":2352,"usd":0.04397,"stage2_stop_reason":"end_turn"},"total_usd":0.05703,"stage1_batch_id":"msgbatch_01DbiymDmubkPpYvdbAu5a7V","stage2_batch_id":"msgbatch_012TbrWvMRH129T7PQ4SmvrJ","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2020,\n      \"finding\": \"AP-2μ (the μ subunit of the AP-2 adaptin complex in P. falciparum) is essential for blood-stage replication: conditional gene knockout severely disrupted schizont organization and maturation and caused mislocalization of key merozoite proteins.\",\n      \"method\": \"Conditional gene knockout with fluorescence microscopy phenotypic readout\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional KO with defined cellular phenotype (schizont disruption, merozoite protein mislocalization), single lab but multiple orthogonal methods (KO + fluorescence + electron microscopy)\",\n      \"pmids\": [\"32098816\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"HA-tagged AP-2μ localizes in P. falciparum as determined by fluorescence and electron microscopy, placing it in a cellular compartment consistent with a trafficking/vesicular function.\",\n      \"method\": \"Fluorescence microscopy and electron microscopy of HA-tagged AP-2μ transgenic parasites\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — direct localization by two imaging modalities in a single lab, but no functional consequence specifically linked to localization site\",\n      \"pmids\": [\"32098816\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Co-immunoprecipitation and mass spectrometry identified AP-2μ-interacting partners including other AP-2 subunits (consistent with a heterotetrameric AP-2 complex), the K10 kelch-domain protein, and PfEHD (an effector of endocytosis and lipid mobilization).\",\n      \"method\": \"Co-immunoprecipitation of HA-tagged AP-2μ followed by mass spectrometry\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP with MS identification, single lab, multiple interactors identified\",\n      \"pmids\": [\"32098816\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"No interaction between AP-2 complex subunits and clathrin was detected; reverse immunoprecipitation with a clathrin nanobody pulled down other heterotetrameric AP-complexes but not AP-2 subunits, indicating the P. falciparum AP-2 complex operates independently of clathrin.\",\n      \"method\": \"Reverse immunoprecipitation using a clathrin nanobody\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — negative result established by reciprocal Co-IP with nanobody, single lab; negative finding is mechanistically informative\",\n      \"pmids\": [\"32098816\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    }\n  ],\n  \"current_model\": \"In P. falciparum, the AP-2 complex β subunit (PF3D7_0528100) is part of a heterotetrameric AP-2 adaptin complex that is essential for schizont maturation and merozoite protein localization during blood-stage replication; the complex interacts with other AP-2 subunits, the K10 kelch-domain protein, and PfEHD, but—unlike canonical AP-2 in other organisms—operates independently of clathrin, indicating a specialized, non-classical endocytic or trafficking function in malaria parasites.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"PF3D7_0528100 is annotated as the putative β subunit of the P. falciparum AP-1/2 adaptin complex, but the available corpus characterizes it only through its membership in the heterotetrameric AP-2 complex rather than through experiments performed on the β subunit itself. The functional studies in this corpus were carried out on the AP-2 μ subunit, whose conditional knockout disrupts schizont organization and maturation and mislocalizes merozoite proteins, establishing the AP-2 complex as essential for blood-stage replication [#0]. Co-immunoprecipitation and mass spectrometry of tagged AP-2μ recovered other AP-2 subunits—consistent with an assembled heterotetramer that would include the β subunit—along with the K10 kelch-domain protein and the endocytic effector PfEHD [#2], and localization imaging places the complex in a compartment consistent with a vesicular/trafficking role [#1]. Notably, reverse immunoprecipitation with a clathrin nanobody failed to recover AP-2 subunits while pulling down other AP complexes, indicating that the P. falciparum AP-2 complex operates independently of clathrin, unlike canonical AP-2 elsewhere [#3]. Beyond its inferred membership in this complex, no experiment in the available corpus directly assays PF3D7_0528100 itself—its individual function, essentiality, and interactions have not been characterized.\",\n  \"teleology\": [\n    {\n      \"year\": 2020,\n      \"claim\": \"Whether the P. falciparum AP-2 complex is required for blood-stage growth was unknown; conditional knockout of the AP-2 μ subunit showed the complex is essential for schizont maturation and correct merozoite protein localization, placing AP-2 in the replicative program.\",\n      \"evidence\": \"Conditional gene knockout of AP-2μ with fluorescence and electron microscopy phenotyping\",\n      \"pmids\": [\"32098816\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"The β subunit PF3D7_0528100 itself was not knocked out; essentiality is demonstrated only for the μ subunit\",\n        \"The precise trafficking cargo and mechanistic step disrupted are not defined\",\n        \"No individual functional assay of PF3D7_0528100\"\n      ],\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"The subcellular context of the AP-2 complex was undefined; imaging of tagged AP-2μ localized it to a compartment consistent with vesicular trafficking, supporting a membrane-associated adaptor role.\",\n      \"evidence\": \"Fluorescence and electron microscopy of HA-tagged AP-2μ transgenic parasites\",\n      \"pmids\": [\"32098816\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Localization was shown for the μ subunit, not for PF3D7_0528100\",\n        \"No functional consequence was tied to the specific localization site\"\n      ],\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"The composition and partners of the complex were unknown; Co-IP/MS of AP-2μ recovered other AP-2 subunits (implying the β subunit assembles into the heterotetramer), the K10 kelch protein, and PfEHD, defining an interaction network for the complex.\",\n      \"evidence\": \"Co-immunoprecipitation of HA-tagged AP-2μ followed by mass spectrometry\",\n      \"pmids\": [\"32098816\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"PF3D7_0528100 is inferred as a complex member but not independently verified by reciprocal pulldown from the β subunit\",\n        \"Functional roles of the K10 and PfEHD interactions are not established\",\n        \"Single lab, single Co-IP bait\"\n      ],\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Whether the parasite AP-2 complex uses the canonical clathrin-dependent pathway was untested; reverse Co-IP with a clathrin nanobody recovered other AP complexes but not AP-2, establishing that P. falciparum AP-2 acts independently of clathrin.\",\n      \"evidence\": \"Reverse immunoprecipitation using a clathrin nanobody\",\n      \"pmids\": [\"32098816\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Negative result does not identify what alternative coat or machinery AP-2 uses\",\n        \"Interaction tested via the μ subunit context, not PF3D7_0528100 directly\"\n      ],\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"No experiment has directly assayed PF3D7_0528100 (the β subunit) itself; its individual contribution to complex assembly, cargo selection, and the clathrin-independent trafficking mechanism remains unknown.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No knockout, tagging, or interaction study performed on PF3D7_0528100 specifically\",\n        \"Molecular function of the β subunit is inferred solely from complex membership\",\n        \"The nature of the clathrin-independent trafficking pathway is undefined\"\n      ],\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [],\n    \"localization\": [\n      {\"term_id\": \"GO:0030136\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0016192\", \"supporting_discovery_ids\": [0, 2, 3]}\n    ],\n    \"complexes\": [\"AP-2 adaptor complex (heterotetrameric)\"],\n    \"partners\": [\"AP-2 complex subunits\", \"K10 kelch-domain protein\", \"PfEHD\"],\n    \"other_free_text\": [\n      \"PF3D7_0528100 is characterized in the corpus only as an inferred β-subunit member of the P. falciparum AP-2 complex; all functional, localization, and interaction data derive from experiments on the AP-2 μ subunit, not the β subunit itself.\",\n      \"The P. falciparum AP-2 complex operates independently of clathrin [#3], distinguishing it from canonical AP-2 adaptors.\",\n      \"AP-2 complex essentiality was demonstrated for schizont maturation and merozoite protein localization during blood-stage replication [#0].\"\n    ]\n  }\n}","audit_flag":{"gene":"PF3D7_0528100","tier":"IDENTITY_CHECK","verdict":"identity unverified","subtype":"names_other_cards","uniprot_band":"","rules_fired":"R12","issue":"No cited paper names this card; 1 of 1 cited papers name other catalogue genes (PF3D7_0304200). The summary may describe one of those genes."},"strains":[{"strain":"Pf3D7","gene_id":"PF3D7_0528100","name":null,"product":"AP-1/2 complex subunit beta, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PbANKA","gene_id":"PBANKA_1242700","name":null,"product":"AP-1/2 complex subunit beta, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PkH","gene_id":"PKNH_1004600","name":null,"product":"AP-1/2 complex subunit beta, putative","uniprot_acc":null,"chromosome":null,"start_pos":null}],"gene_symbol":"","evaluation":null}