{"gene":"PF3D7_1422900","run_date":"2026-09-08T14:19:07","timeline":{"discoveries":[{"year":2016,"finding":"Pf14-3-3I (PF3D7_1422900) physically interacts with the AKAP-like protein PfAKAL, as demonstrated by pull-down experiments using recombinant proteins and parasite protein extracts, and the two proteins colocalize in P. falciparum.","method":"Pull-down assay (recombinant protein and parasite extracts), immunofluorescence colocalization","journal":"Malaria journal","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single lab, reciprocal pull-down with recombinant proteins plus colocalization, two orthogonal methods","pmids":["27129434"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"finding":"Pf14-3-3I (PF3D7_1422900) interacts with the PKA regulatory subunit PfPKA-R, suggesting it bridges PfAKAL to PKA rather than PfAKAL binding PKA-R directly.","method":"Pull-down assay and biochemical/proteomic interaction analysis","journal":"Malaria journal","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single lab, pull-down plus proteomic confirmation, two orthogonal methods","pmids":["27129434"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"finding":"Pf14-3-3I (PF3D7_1422900) is phosphorylated in vitro by PKA (whereas PfAKAL is not), indicating PKA-mediated phosphorylation as a regulatory mechanism for Pf14-3-3I-containing complexes.","method":"Radiolabel kinase activity assay (in vitro phosphorylation)","journal":"Malaria journal","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — in vitro kinase assay with a defined writer (PKA), single lab, single method","pmids":["27129434"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]}],"current_model":"In P. falciparum, Pf14-3-3I (PF3D7_1422900) physically associates with the AKAP-like protein PfAKAL and with the PKA regulatory subunit PfPKA-R, is itself phosphorylated by PKA in vitro, and thereby appears to act as an indirect bridge linking PfAKAL to the cAMP/PKA signaling pathway during asexual intra-erythrocytic development."},"narrative":{"mechanistic_narrative":"Pf14-3-3I (PF3D7_1422900) is a P. falciparum 14-3-3 protein that functions as a scaffolding adaptor coupling an AKAP-like protein to the cAMP/PKA signaling machinery in the asexual blood stage [PMID:27129434]. In P. falciparum 3D7, it physically associates with the AKAP-like protein PfAKAL, with which it colocalizes in the parasite [PMID:27129434], and it independently binds the PKA regulatory subunit PfPKA-R, positioning it as an indirect bridge that links PfAKAL to PKA rather than PfAKAL contacting PKA-R directly [PMID:27129434]. Pf14-3-3I is itself a PKA substrate, being phosphorylated in vitro by PKA whereas PfAKAL is not, implying that PKA-mediated phosphorylation regulates Pf14-3-3I-containing complexes [PMID:27129434]. Beyond these interaction and phosphorylation findings, no further mechanistic detail — including the functional consequence of the interactions in vivo, the phosphosite involved, or any essentiality data for PF3D7_1422900 — has been characterized in the available corpus.","teleology":[{"year":2016,"claim":"Establishing that Pf14-3-3I directly binds the AKAP-like protein PfAKAL identified the first molecular partner of this 14-3-3 protein and placed it within an anchoring-protein context.","evidence":"Reciprocal pull-down with recombinant proteins and parasite extracts plus immunofluorescence colocalization in P. falciparum 3D7","pmids":["27129434"],"confidence":"Medium","gaps":["Does not define the binding interface or whether the interaction is phosphorylation-dependent","Functional consequence of the PfAKAL association in vivo is not established","Colocalization does not resolve the subcellular compartment"],"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"claim":"Demonstrating that Pf14-3-3I also binds the PKA regulatory subunit PfPKA-R answered how PfAKAL connects to PKA, casting Pf14-3-3I as an indirect bridge rather than PfAKAL binding PKA-R directly.","evidence":"Pull-down assay with biochemical/proteomic interaction analysis in P. falciparum 3D7","pmids":["27129434"],"confidence":"Medium","gaps":["Ternary complex of PfAKAL–Pf14-3-3I–PfPKA-R not directly reconstituted","Whether the two interactions are simultaneous or mutually exclusive is unresolved","In vivo relevance to cAMP/PKA signaling not demonstrated"],"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"claim":"Showing Pf14-3-3I is phosphorylated by PKA in vitro (while PfAKAL is not) identified a candidate regulatory mechanism controlling Pf14-3-3I-containing complexes.","evidence":"Radiolabel in vitro kinase activity assay with PKA as the writer","pmids":["27129434"],"confidence":"Medium","gaps":["In vitro kinase assay does not confirm phosphorylation occurs in the parasite","Phosphosite(s) not mapped","Effect of phosphorylation on PfAKAL or PfPKA-R binding not tested"],"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":null,"claim":"The physiological role of Pf14-3-3I in asexual development and the functional output of its bridging of PfAKAL to PKA remain unknown.","evidence":"No genetic loss-of-function, phenotypic, or structural data for PF3D7_1422900 in the corpus","pmids":[],"confidence":"Low","gaps":["No knockout or knockdown phenotype for PF3D7_1422900","No structure of the complex or defined binding motifs","No demonstration that PKA phosphorylation of Pf14-3-3I alters complex assembly or signaling in vivo"],"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["PfAKAL–Pf14-3-3I–PfPKA-R (cAMP/PKA anchoring complex, inferred from pairwise interactions)"],"partners":["PFAKAL (AKAP-LIKE PROTEIN)","PFPKA-R (PKA REGULATORY SUBUNIT)"],"other_free_text":["Pf14-3-3I is a substrate of PKA in vitro; PfAKAL is not phosphorylated by PKA [#2]","All findings derive from a single study in P. falciparum 3D7 (PMID 27129434)"]}},"prefetch_data":{},"corpus_meta":[{"pmid":"22984113","id":"PMC_22984113","title":"High levels of immunoglobulin E autoantibody to 14-3-3 epsilon protein correlate with protection against severe Plasmodium falciparum malaria.","date":"2012","source":"The Journal of infectious diseases","url":"https://pubmed.ncbi.nlm.nih.gov/22984113","citation_count":14,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["14-3-3 protein"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"27129434","id":"PMC_27129434","title":"Characterization of an A-kinase anchoring protein-like suggests an alternative way of PKA anchoring in Plasmodium falciparum.","date":"2016","source":"Malaria journal","url":"https://pubmed.ncbi.nlm.nih.gov/27129434","citation_count":6,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["14-3-3 protein"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":4195,"output_tokens":812,"usd":0.012382,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":5406,"output_tokens":1983,"usd":0.038303,"stage2_stop_reason":"end_turn"},"total_usd":0.050685,"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\": 2016,\n      \"finding\": \"Pf14-3-3I (PF3D7_1422900) physically interacts with the AKAP-like protein PfAKAL, as demonstrated by pull-down experiments using recombinant proteins and parasite protein extracts, and the two proteins colocalize in P. falciparum.\",\n      \"method\": \"Pull-down assay (recombinant protein and parasite extracts), immunofluorescence colocalization\",\n      \"journal\": \"Malaria journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single lab, reciprocal pull-down with recombinant proteins plus colocalization, two orthogonal methods\",\n      \"pmids\": [\"27129434\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Pf14-3-3I (PF3D7_1422900) interacts with the PKA regulatory subunit PfPKA-R, suggesting it bridges PfAKAL to PKA rather than PfAKAL binding PKA-R directly.\",\n      \"method\": \"Pull-down assay and biochemical/proteomic interaction analysis\",\n      \"journal\": \"Malaria journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single lab, pull-down plus proteomic confirmation, two orthogonal methods\",\n      \"pmids\": [\"27129434\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Pf14-3-3I (PF3D7_1422900) is phosphorylated in vitro by PKA (whereas PfAKAL is not), indicating PKA-mediated phosphorylation as a regulatory mechanism for Pf14-3-3I-containing complexes.\",\n      \"method\": \"Radiolabel kinase activity assay (in vitro phosphorylation)\",\n      \"journal\": \"Malaria journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — in vitro kinase assay with a defined writer (PKA), single lab, single method\",\n      \"pmids\": [\"27129434\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    }\n  ],\n  \"current_model\": \"In P. falciparum, Pf14-3-3I (PF3D7_1422900) physically associates with the AKAP-like protein PfAKAL and with the PKA regulatory subunit PfPKA-R, is itself phosphorylated by PKA in vitro, and thereby appears to act as an indirect bridge linking PfAKAL to the cAMP/PKA signaling pathway during asexual intra-erythrocytic development.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"Pf14-3-3I (PF3D7_1422900) is a P. falciparum 14-3-3 protein that functions as a scaffolding adaptor coupling an AKAP-like protein to the cAMP/PKA signaling machinery in the asexual blood stage [#0, #1]. In P. falciparum 3D7, it physically associates with the AKAP-like protein PfAKAL, with which it colocalizes in the parasite [#0], and it independently binds the PKA regulatory subunit PfPKA-R, positioning it as an indirect bridge that links PfAKAL to PKA rather than PfAKAL contacting PKA-R directly [#1]. Pf14-3-3I is itself a PKA substrate, being phosphorylated in vitro by PKA whereas PfAKAL is not, implying that PKA-mediated phosphorylation regulates Pf14-3-3I-containing complexes [#2]. Beyond these interaction and phosphorylation findings, no further mechanistic detail — including the functional consequence of the interactions in vivo, the phosphosite involved, or any essentiality data for PF3D7_1422900 — has been characterized in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 2016,\n      \"claim\": \"Establishing that Pf14-3-3I directly binds the AKAP-like protein PfAKAL identified the first molecular partner of this 14-3-3 protein and placed it within an anchoring-protein context.\",\n      \"evidence\": \"Reciprocal pull-down with recombinant proteins and parasite extracts plus immunofluorescence colocalization in P. falciparum 3D7\",\n      \"pmids\": [\"27129434\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Does not define the binding interface or whether the interaction is phosphorylation-dependent\",\n        \"Functional consequence of the PfAKAL association in vivo is not established\",\n        \"Colocalization does not resolve the subcellular compartment\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrating that Pf14-3-3I also binds the PKA regulatory subunit PfPKA-R answered how PfAKAL connects to PKA, casting Pf14-3-3I as an indirect bridge rather than PfAKAL binding PKA-R directly.\",\n      \"evidence\": \"Pull-down assay with biochemical/proteomic interaction analysis in P. falciparum 3D7\",\n      \"pmids\": [\"27129434\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Ternary complex of PfAKAL–Pf14-3-3I–PfPKA-R not directly reconstituted\",\n        \"Whether the two interactions are simultaneous or mutually exclusive is unresolved\",\n        \"In vivo relevance to cAMP/PKA signaling not demonstrated\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Showing Pf14-3-3I is phosphorylated by PKA in vitro (while PfAKAL is not) identified a candidate regulatory mechanism controlling Pf14-3-3I-containing complexes.\",\n      \"evidence\": \"Radiolabel in vitro kinase activity assay with PKA as the writer\",\n      \"pmids\": [\"27129434\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"In vitro kinase assay does not confirm phosphorylation occurs in the parasite\",\n        \"Phosphosite(s) not mapped\",\n        \"Effect of phosphorylation on PfAKAL or PfPKA-R binding not tested\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The physiological role of Pf14-3-3I in asexual development and the functional output of its bridging of PfAKAL to PKA remain unknown.\",\n      \"evidence\": \"No genetic loss-of-function, phenotypic, or structural data for PF3D7_1422900 in the corpus\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No knockout or knockdown phenotype for PF3D7_1422900\",\n        \"No structure of the complex or defined binding motifs\",\n        \"No demonstration that PKA phosphorylation of Pf14-3-3I alters complex assembly or signaling in vivo\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0019904\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0030552\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"GO:0034199\", \"supporting_discovery_ids\": [1, 2]}\n    ],\n    \"complexes\": [\n      \"PfAKAL–Pf14-3-3I–PfPKA-R (cAMP/PKA anchoring complex, inferred from pairwise interactions)\"\n    ],\n    \"partners\": [\n      \"PfAKAL (AKAP-like protein)\",\n      \"PfPKA-R (PKA regulatory subunit)\"\n    ],\n    \"other_free_text\": [\n      \"Pf14-3-3I is a substrate of PKA in vitro; PfAKAL is not phosphorylated by PKA [#2]\",\n      \"All findings derive from a single study in P. falciparum 3D7 (PMID 27129434)\"\n    ]\n  }\n}","audit_flag":{"gene":"PF3D7_1422900","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_0512900, PF3D7_0818200, PF3D7_0934800). The summary may describe one of those genes."},"strains":[{"strain":"Pf3D7","gene_id":"PF3D7_1422900","name":null,"product":"14-3-3 protein, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PbANKA","gene_id":"PBANKA_1020500","name":null,"product":"14-3-3 protein, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PkH","gene_id":"PKNH_1335100","name":null,"product":"14-3-3 protein, putative","uniprot_acc":null,"chromosome":null,"start_pos":null}],"gene_symbol":"","evaluation":null}