{"gene":"PF3D7_0414600","run_date":"2026-09-08T19:44:17","timeline":{"discoveries":[{"year":2018,"finding":"APH (PF3D7_0414600) contains a pleckstrin-homology domain that directly senses phosphatidic acid (PA); crystal structure of TgAPH resolved the atomic details of the PA-sensing hub, showing that APH inserts into lipid bilayers and clusters multiple phosphate head-groups at the bilayer binding surface. The key determinants of PA recognition were identified by structural and functional dissection.","method":"Crystal structure determination combined with functional interaction studies with phospholipid membranes (structural biology + lipid-binding assays)","journal":"Structure (London, England : 1993)","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure with functional validation of PA-binding mechanism, published in peer-reviewed journal; although performed primarily in TgAPH, the paper explicitly addresses PfAPH in the context of the conserved PA-sensing hub","pmids":["29910186"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""},{"species":"Toxoplasma gondii","strain":""}]},{"year":2021,"finding":"APH (PF3D7_0414600) is required for microneme exocytosis in Plasmodium yoelii; inducible deletion of APH impaired secretion of the microneme proteins MTRAP and AMA1, reduced asexual growth and erythrocyte invasion, and reduced echinocytosis after merozoite internalization. APH largely colocalized with MTRAP but less with AMA1, indicating a direct role in MTRAP secretion and only an indirect role in AMA1 secretion.","method":"DiCre-based inducible knockout in P. yoelii with secretion assays (MTRAP, AMA1), invasion assays, IFA colocalization, and electron microscopy of tight junction formation","journal":"Parasitology international","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean inducible KO with multiple orthogonal phenotypic readouts (secretion, invasion, echinocytosis, colocalization) in a single rigorous study","pmids":["34628068"],"is_preprint":false,"attribution":[{"species":"Plasmodium yoelii","strain":""}]},{"year":2021,"finding":"APH (PF3D7_0414600) undergoes N-myristoylation at its N-terminal glycine in P. falciparum; the modified N-terminal peptide was directly detected by chemical proteomics using the alkyne-myristate probe YnMyr and NMT inhibitor treatment, confirming APH as a substrate of N-myristoyl transferase (NMT).","method":"Chemical proteomics: metabolic labeling with alkyne-myristate (YnMyr) followed by enrichment and MS/MS identification of the modified N-terminal peptide; NMT inhibitor (IMP-1002) treatment as control","journal":"PLoS biology","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — direct biochemical identification of the modification by MS with diagnostic b1 ion, but single study, single method for this specific protein","pmids":["34695132"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2019,"finding":"APH (PF3D7_0414600) localizes to the surface of micronemes and its PA-binding activity initiates membrane fusion for microneme exocytosis at the apical end of the parasite plasma membrane, where PA is generated prior to egress. This places APH upstream of rhoptry discharge in the sequential secretion pathway.","method":"Referenced as established prior work (cited as reference 27 in the RASP2 study); the RASP2 paper uses APH localization and function as a mechanistic comparator/framework","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — localization and functional role cited as established across multiple papers; the specific mechanistic claim is supported by convergent evidence from PMID 29910186 and 34628068, though this paper itself only cites prior work on APH","pmids":["31492901"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""},{"species":"Toxoplasma gondii","strain":""}]},{"year":2025,"finding":"APH (PF3D7_0414600) localizes apically in P. falciparum schizonts independently of HOPS/CORVET function; when HOPS/CORVET core subunits (VPS16, VPS18) were depleted, APH-mCherry retained its typical apical localization, indicating that microneme biogenesis (as marked by APH surface anchoring) is not disrupted by HOPS/CORVET depletion, even though secreted microneme cargo proteins were mislocalized.","method":"Conditional knockdown of HOPS/CORVET subunits combined with live fluorescence microscopy of APH-mCherry as a microneme surface/biogenesis marker","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean KO/KD with direct localization imaging, single study, used APH as a validated organelle marker with a specific negative result that is mechanistically informative (HOPS/CORVET not required for microneme biogenesis)","pmids":["40198740"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"current_model":"APH (PF3D7_0414600) is an N-myristoylated and palmitoylated protein anchored on the outer surface of micronemes that directly senses phosphatidic acid (PA) through its pleckstrin-homology domain — inserting into and clustering PA head-groups at the bilayer — thereby providing lipid-dependent spatial control for microneme docking and fusion at the PA-enriched apical plasma membrane, with direct involvement in MTRAP secretion and an indirect role in AMA1 secretion during erythrocyte invasion."},"narrative":{"mechanistic_narrative":"APH (PF3D7_0414600) is a lipid-sensing protein anchored to the surface of micronemes that provides spatial control for microneme exocytosis during apicomplexan host-cell invasion [PMID:29910186, PMID:31492901]. Its pleckstrin-homology domain directly senses phosphatidic acid (PA): structural and functional dissection showed that APH inserts into lipid bilayers and clusters multiple PA head-groups at its binding surface, defining a conserved PA-sensing hub [PMID:29910186]. Surface anchoring is achieved through N-terminal lipid modification, with APH confirmed as an N-myristoyltransferase substrate that is myristoylated at its N-terminal glycine in P. falciparum [PMID:34695132]. Functionally, inducible deletion in P. yoelii established that APH is required for microneme exocytosis, erythrocyte invasion, and asexual growth: APH loss impaired secretion of the microneme protein MTRAP and reduced post-internalization echinocytosis, with APH colocalizing largely with MTRAP and less with AMA1, indicating a direct role in MTRAP secretion and only an indirect role in AMA1 secretion [PMID:34628068]. APH marks micronemes independently of the HOPS/CORVET tethering machinery, as its apical localization persists when VPS16/VPS18 are depleted despite mislocalization of secreted microneme cargo [PMID:40198740]. Beyond PA-sensing and its role positioning microneme fusion at the apical membrane, the corpus does not characterize additional binding partners or downstream fusion machinery for APH.","teleology":[{"year":2018,"claim":"Establishing how APH engages membranes answered whether the protein is a genuine lipid sensor, revealing a PA-recognition mechanism rather than a generic membrane anchor.","evidence":"Crystal structure of the APH PH domain with phospholipid-binding assays (largely in T. gondii, addressing conserved PfAPH hub)","pmids":["29910186"],"confidence":"High","gaps":["Structural work performed primarily on TgAPH, not the P. falciparum protein directly","Does not establish the in vivo functional consequence of PA sensing","Downstream fusion partners engaged after PA binding not identified"],"attribution":[{"species":"Plasmodium falciparum","strain":""},{"species":"Toxoplasma gondii","strain":""}]},{"year":2019,"claim":"Placing APH in the secretion sequence framed its PA-binding activity as the trigger initiating microneme membrane fusion at the apical plasma membrane, upstream of rhoptry discharge.","evidence":"APH localization and function cited as a mechanistic comparator framework within the RASP2 study","pmids":["31492901"],"confidence":"Medium","gaps":["This paper cites prior APH work rather than generating new APH data","Temporal ordering relative to rhoptry discharge inferred, not directly measured for APH","PA generation source at the apical membrane not defined here"],"attribution":[{"species":"Plasmodium falciparum","strain":""},{"species":"Toxoplasma gondii","strain":""}]},{"year":2021,"claim":"Inducible deletion answered whether APH is functionally required for invasion, demonstrating it is needed for microneme exocytosis and defining a direct role in MTRAP versus an indirect role in AMA1 secretion.","evidence":"DiCre inducible knockout in P. yoelii with secretion, invasion, echinocytosis, colocalization, and electron microscopy readouts","pmids":["34628068"],"confidence":"High","gaps":["Molecular basis for differential MTRAP vs AMA1 dependence not resolved","Direct physical interaction between APH and MTRAP not shown","Performed in P. yoelii; not confirmed by equivalent P. falciparum knockout"],"attribution":[{"species":"Plasmodium yoelii","strain":""}]},{"year":2021,"claim":"Identifying the N-terminal modification answered how APH is targeted to the microneme surface, confirming N-myristoylation as the lipid anchor and APH as an NMT substrate.","evidence":"Chemical proteomics with YnMyr metabolic labeling, enrichment, and MS/MS of the modified N-terminal peptide, with NMT inhibitor control in P. falciparum","pmids":["34695132"],"confidence":"Medium","gaps":["Single method for this specific protein","Whether myristoylation alone or combined with palmitoylation drives stable membrane anchoring not tested","Functional consequence of blocking APH myristoylation not directly measured"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2025,"claim":"Testing APH localization under HOPS/CORVET depletion distinguished microneme biogenesis/surface anchoring from cargo delivery, showing APH surface anchoring is independent of the tethering complex even when secreted cargo is mislocalized.","evidence":"Conditional knockdown of VPS16/VPS18 with live imaging of APH-mCherry as a microneme surface marker in P. falciparum","pmids":["40198740"],"confidence":"Medium","gaps":["APH used as a marker rather than the primary subject of perturbation","Machinery that delivers APH itself to the microneme surface not identified","Does not resolve how cargo mislocalization arises independently of APH anchoring"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":null,"claim":"How APH couples PA sensing to the downstream membrane-fusion machinery and to selective MTRAP secretion remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No direct APH binding partners or fusion effectors identified in the corpus","Mechanistic basis for direct MTRAP versus indirect AMA1 dependence unknown","P. falciparum-specific functional knockout phenotype not established"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":[],"partners":["MTRAP (COLOCALIZES WITH APH; APH REQUIRED FOR ITS SECRETION) [#1]","N-MYRISTOYLTRANSFERASE (NMT) — MODIFYING ENZYME [#2]"],"other_free_text":["APH is N-myristoylated at its N-terminal glycine, consistent with surface membrane anchoring on micronemes [#2].","APH-mCherry serves as a validated microneme surface/biogenesis marker; its apical localization is independent of HOPS/CORVET (VPS16/VPS18) function [#4].","APH loss reduces asexual growth, erythrocyte invasion, and post-internalization echinocytosis in P. yoelii [#1]."]}},"prefetch_data":{},"corpus_meta":[{"pmid":"29910186","id":"PMC_29910186","title":"Structural Basis of Phosphatidic Acid Sensing by APH in Apicomplexan Parasites.","date":"2018","source":"Structure (London, England : 1993)","url":"https://pubmed.ncbi.nlm.nih.gov/29910186","citation_count":17,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["APH"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"34628068","id":"PMC_34628068","title":"Distinct effects on the secretion of MTRAP and AMA1 in Plasmodium yoelii following deletion of acylated pleckstrin homology domain-containing protein.","date":"2021","source":"Parasitology international","url":"https://pubmed.ncbi.nlm.nih.gov/34628068","citation_count":2,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["APH"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"42211658","id":"","title":"Physiology of invasive Plasmodium merozoites.","date":"2026","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/42211658","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["APH"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"42233156","id":"","title":"A pleckstrin homology domain protein is involved in crystalloid formation in Plasmodium ookinetes and affects the maturation of infective sporozoites.","date":"2026","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/42233156","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["APH"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"40198740","id":"","title":"HOPS/CORVET tethering complexes are critical for endocytosis and protein trafficking to invasion related organelles in malaria parasites.","date":"2025","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/40198740","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["APH"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"34695132","id":"","title":"Inhibition of protein N-myristoylation blocks Plasmodium falciparum intraerythrocytic development, egress and invasion.","date":"2021","source":"PLoS biology","url":"https://pubmed.ncbi.nlm.nih.gov/34695132","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["APH"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"36622145","id":"","title":"Targetome Analysis of Malaria Sporozoite Transcription Factor AP2-Sp Reveals Its Role as a Master Regulator.","date":"2023","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/36622145","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["APH"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"33082261","id":"","title":"The ZIP Code of Vesicle Trafficking in Apicomplexa: SEC1/Munc18 and SNARE Proteins.","date":"2020","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/33082261","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["APH"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"30111801","id":"","title":"Unraveling the Plasmodium vivax sporozoite transcriptional journey from mosquito vector to human host.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/30111801","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["APH"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"31492901","id":"","title":"A lipid-binding protein mediates rhoptry discharge and invasion in Plasmodium falciparum and Toxoplasma gondii parasites.","date":"2019","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/31492901","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["APH"],"competing_organism_flag":true,"competing_organism_match":"Toxoplasma","competing_organism_only":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":13570,"output_tokens":1723,"usd":0.033278,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6763,"output_tokens":2774,"usd":0.051583,"stage2_stop_reason":"end_turn"},"total_usd":0.084861,"stage1_batch_id":"msgbatch_01SRp5hyRc82zNjddxyCCXGk","stage2_batch_id":"msgbatch_01K3BTmBazRo3pRrMvyaPMTY","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2018,\n      \"finding\": \"APH (PF3D7_0414600) contains a pleckstrin-homology domain that directly senses phosphatidic acid (PA); crystal structure of TgAPH resolved the atomic details of the PA-sensing hub, showing that APH inserts into lipid bilayers and clusters multiple phosphate head-groups at the bilayer binding surface. The key determinants of PA recognition were identified by structural and functional dissection.\",\n      \"method\": \"Crystal structure determination combined with functional interaction studies with phospholipid membranes (structural biology + lipid-binding assays)\",\n      \"journal\": \"Structure (London, England : 1993)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure with functional validation of PA-binding mechanism, published in peer-reviewed journal; although performed primarily in TgAPH, the paper explicitly addresses PfAPH in the context of the conserved PA-sensing hub\",\n      \"pmids\": [\"29910186\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"},\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"APH (PF3D7_0414600) is required for microneme exocytosis in Plasmodium yoelii; inducible deletion of APH impaired secretion of the microneme proteins MTRAP and AMA1, reduced asexual growth and erythrocyte invasion, and reduced echinocytosis after merozoite internalization. APH largely colocalized with MTRAP but less with AMA1, indicating a direct role in MTRAP secretion and only an indirect role in AMA1 secretion.\",\n      \"method\": \"DiCre-based inducible knockout in P. yoelii with secretion assays (MTRAP, AMA1), invasion assays, IFA colocalization, and electron microscopy of tight junction formation\",\n      \"journal\": \"Parasitology international\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean inducible KO with multiple orthogonal phenotypic readouts (secretion, invasion, echinocytosis, colocalization) in a single rigorous study\",\n      \"pmids\": [\"34628068\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Plasmodium yoelii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"APH (PF3D7_0414600) undergoes N-myristoylation at its N-terminal glycine in P. falciparum; the modified N-terminal peptide was directly detected by chemical proteomics using the alkyne-myristate probe YnMyr and NMT inhibitor treatment, confirming APH as a substrate of N-myristoyl transferase (NMT).\",\n      \"method\": \"Chemical proteomics: metabolic labeling with alkyne-myristate (YnMyr) followed by enrichment and MS/MS identification of the modified N-terminal peptide; NMT inhibitor (IMP-1002) treatment as control\",\n      \"journal\": \"PLoS biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — direct biochemical identification of the modification by MS with diagnostic b1 ion, but single study, single method for this specific protein\",\n      \"pmids\": [\"34695132\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"APH (PF3D7_0414600) localizes to the surface of micronemes and its PA-binding activity initiates membrane fusion for microneme exocytosis at the apical end of the parasite plasma membrane, where PA is generated prior to egress. This places APH upstream of rhoptry discharge in the sequential secretion pathway.\",\n      \"method\": \"Referenced as established prior work (cited as reference 27 in the RASP2 study); the RASP2 paper uses APH localization and function as a mechanistic comparator/framework\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — localization and functional role cited as established across multiple papers; the specific mechanistic claim is supported by convergent evidence from PMID 29910186 and 34628068, though this paper itself only cites prior work on APH\",\n      \"pmids\": [\"31492901\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"},\n        {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"APH (PF3D7_0414600) localizes apically in P. falciparum schizonts independently of HOPS/CORVET function; when HOPS/CORVET core subunits (VPS16, VPS18) were depleted, APH-mCherry retained its typical apical localization, indicating that microneme biogenesis (as marked by APH surface anchoring) is not disrupted by HOPS/CORVET depletion, even though secreted microneme cargo proteins were mislocalized.\",\n      \"method\": \"Conditional knockdown of HOPS/CORVET subunits combined with live fluorescence microscopy of APH-mCherry as a microneme surface/biogenesis marker\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean KO/KD with direct localization imaging, single study, used APH as a validated organelle marker with a specific negative result that is mechanistically informative (HOPS/CORVET not required for microneme biogenesis)\",\n      \"pmids\": [\"40198740\"],\n      \"is_preprint\": false,\n      \"attribution\": [\n        {\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}\n      ]\n    }\n  ],\n  \"current_model\": \"APH (PF3D7_0414600) is an N-myristoylated and palmitoylated protein anchored on the outer surface of micronemes that directly senses phosphatidic acid (PA) through its pleckstrin-homology domain — inserting into and clustering PA head-groups at the bilayer — thereby providing lipid-dependent spatial control for microneme docking and fusion at the PA-enriched apical plasma membrane, with direct involvement in MTRAP secretion and an indirect role in AMA1 secretion during erythrocyte invasion.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"APH (PF3D7_0414600) is a lipid-sensing protein anchored to the surface of micronemes that provides spatial control for microneme exocytosis during apicomplexan host-cell invasion [#0, #3]. Its pleckstrin-homology domain directly senses phosphatidic acid (PA): structural and functional dissection showed that APH inserts into lipid bilayers and clusters multiple PA head-groups at its binding surface, defining a conserved PA-sensing hub [#0]. Surface anchoring is achieved through N-terminal lipid modification, with APH confirmed as an N-myristoyltransferase substrate that is myristoylated at its N-terminal glycine in P. falciparum [#2]. Functionally, inducible deletion in P. yoelii established that APH is required for microneme exocytosis, erythrocyte invasion, and asexual growth: APH loss impaired secretion of the microneme protein MTRAP and reduced post-internalization echinocytosis, with APH colocalizing largely with MTRAP and less with AMA1, indicating a direct role in MTRAP secretion and only an indirect role in AMA1 secretion [#1]. APH marks micronemes independently of the HOPS/CORVET tethering machinery, as its apical localization persists when VPS16/VPS18 are depleted despite mislocalization of secreted microneme cargo [#4]. Beyond PA-sensing and its role positioning microneme fusion at the apical membrane, the corpus does not characterize additional binding partners or downstream fusion machinery for APH.\",\n  \"teleology\": [\n    {\n      \"year\": 2018,\n      \"claim\": \"Establishing how APH engages membranes answered whether the protein is a genuine lipid sensor, revealing a PA-recognition mechanism rather than a generic membrane anchor.\",\n      \"evidence\": \"Crystal structure of the APH PH domain with phospholipid-binding assays (largely in T. gondii, addressing conserved PfAPH hub)\",\n      \"pmids\": [\"29910186\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural work performed primarily on TgAPH, not the P. falciparum protein directly\", \"Does not establish the in vivo functional consequence of PA sensing\", \"Downstream fusion partners engaged after PA binding not identified\"],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}, {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Placing APH in the secretion sequence framed its PA-binding activity as the trigger initiating microneme membrane fusion at the apical plasma membrane, upstream of rhoptry discharge.\",\n      \"evidence\": \"APH localization and function cited as a mechanistic comparator framework within the RASP2 study\",\n      \"pmids\": [\"31492901\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"This paper cites prior APH work rather than generating new APH data\", \"Temporal ordering relative to rhoptry discharge inferred, not directly measured for APH\", \"PA generation source at the apical membrane not defined here\"],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}, {\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Inducible deletion answered whether APH is functionally required for invasion, demonstrating it is needed for microneme exocytosis and defining a direct role in MTRAP versus an indirect role in AMA1 secretion.\",\n      \"evidence\": \"DiCre inducible knockout in P. yoelii with secretion, invasion, echinocytosis, colocalization, and electron microscopy readouts\",\n      \"pmids\": [\"34628068\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis for differential MTRAP vs AMA1 dependence not resolved\", \"Direct physical interaction between APH and MTRAP not shown\", \"Performed in P. yoelii; not confirmed by equivalent P. falciparum knockout\"],\n      \"attribution\": [{\"species\": \"Plasmodium yoelii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identifying the N-terminal modification answered how APH is targeted to the microneme surface, confirming N-myristoylation as the lipid anchor and APH as an NMT substrate.\",\n      \"evidence\": \"Chemical proteomics with YnMyr metabolic labeling, enrichment, and MS/MS of the modified N-terminal peptide, with NMT inhibitor control in P. falciparum\",\n      \"pmids\": [\"34695132\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single method for this specific protein\", \"Whether myristoylation alone or combined with palmitoylation drives stable membrane anchoring not tested\", \"Functional consequence of blocking APH myristoylation not directly measured\"],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Testing APH localization under HOPS/CORVET depletion distinguished microneme biogenesis/surface anchoring from cargo delivery, showing APH surface anchoring is independent of the tethering complex even when secreted cargo is mislocalized.\",\n      \"evidence\": \"Conditional knockdown of VPS16/VPS18 with live imaging of APH-mCherry as a microneme surface marker in P. falciparum\",\n      \"pmids\": [\"40198740\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"APH used as a marker rather than the primary subject of perturbation\", \"Machinery that delivers APH itself to the microneme surface not identified\", \"Does not resolve how cargo mislocalization arises independently of APH anchoring\"],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How APH couples PA sensing to the downstream membrane-fusion machinery and to selective MTRAP secretion remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct APH binding partners or fusion effectors identified in the corpus\", \"Mechanistic basis for direct MTRAP versus indirect AMA1 dependence unknown\", \"P. falciparum-specific functional knockout phenotype not established\"],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0070300\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [0, 3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0020009\", \"supporting_discovery_ids\": [1, 3, 4]},\n      {\"term_id\": \"GO:0020005\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0060631\", \"supporting_discovery_ids\": [1, 3]},\n      {\"term_id\": \"GO:0006887\", \"supporting_discovery_ids\": [1, 3]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"MTRAP (colocalizes with APH; APH required for its secretion) [#1]\",\n      \"N-myristoyltransferase (NMT) — modifying enzyme [#2]\"\n    ],\n    \"other_free_text\": [\n      \"APH is N-myristoylated at its N-terminal glycine, consistent with surface membrane anchoring on micronemes [#2].\",\n      \"APH-mCherry serves as a validated microneme surface/biogenesis marker; its apical localization is independent of HOPS/CORVET (VPS16/VPS18) function [#4].\",\n      \"APH loss reduces asexual growth, erythrocyte invasion, and post-internalization echinocytosis in P. yoelii [#1].\"\n    ]\n  }\n}","audit_flag":null,"strains":[{"strain":"Pf3D7","gene_id":"PF3D7_0414600","name":null,"product":"acylated pleckstrin-homology domain-containing protein, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PbANKA","gene_id":"PBANKA_0716600","name":null,"product":"acylated pleckstrin-homology domain-containing protein, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PkH","gene_id":"PKNH_0506200","name":null,"product":"acylated pleckstrin-homology domain-containing protein, putative","uniprot_acc":null,"chromosome":null,"start_pos":null}],"gene_symbol":"APH","evaluation":null}