{"gene":"PF3D7_0512900","run_date":"2026-09-07T10:55:52","timeline":{"discoveries":[{"year":2016,"finding":"AKAL (PF3D7_0512900) is expressed in mature asexual stages, merozoites, and gametocytes of P. falciparum 3D7, as determined by RT-qPCR and immunofluorescence assays.","method":"RT-qPCR and immunofluorescence assay","journal":"Malaria journal","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — two orthogonal methods (RT-qPCR + IFA) in a single lab, but no functional consequence directly linked to localization","pmids":["27129434"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"finding":"AKAL (PF3D7_0512900) does NOT interact directly with the P. falciparum PKA regulatory subunit (PfPKA-R); instead, AKAL binds Pf14-3-3I, which in turn interacts with PfPKA-R, establishing an indirect connection to PKA signaling.","method":"Pull-down experiments with recombinant AKAL protein and parasite protein extracts, biochemical and proteomic approaches, immunofluorescence colocalization","journal":"Malaria journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal pull-down plus proteomic confirmation and IFA colocalization, single lab","pmids":["27129434"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"finding":"AKAL (PF3D7_0512900) is NOT phosphorylated by PKA in vitro, whereas its binding partner Pf14-3-3I and PfPKA-R are phosphorylated by PKA, indicating that phosphorylation of the complex components (but not AKAL itself) may regulate these interactions.","method":"Radiolabel in vitro kinase activity assay","journal":"Malaria journal","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — in vitro biochemical assay, single lab, single method","pmids":["27129434"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"finding":"AKAL (PF3D7_0512900) binds nucleotides AMP and cAMP, as demonstrated by qualitative nucleotide binding experiments and structural modeling that revealed a conserved nucleotide-binding pocket, suggesting possible involvement in AMPK pathway or phosphodiesterase-associated activities.","method":"Structural modeling and qualitative nucleotide binding experiments","journal":"Malaria journal","confidence":"Low","confidence_rationale":"Tier 3 / Weak — qualitative binding assay supported by computational structural model, single lab, no functional validation","pmids":["27129434"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"finding":"AKAL (PF3D7_0512900) shares 23% sequence identity with the central domain of human AKAP18δ but represents an atypical AKAP in that it does not anchor PKA-R directly; instead PKA localization in P. falciparum may involve other partners.","method":"Sequence alignment and biochemical interaction studies","journal":"Malaria journal","confidence":"Low","confidence_rationale":"Tier 3 / Weak — sequence comparison combined with negative interaction result, single lab","pmids":["27129434"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]}],"current_model":"AKAL (PF3D7_0512900) is an atypical AKAP-like protein expressed in mature asexual stages, merozoites, and gametocytes of P. falciparum that does not directly bind the PKA regulatory subunit but instead interacts with Pf14-3-3I (which itself binds PfPKA-R), is not a PKA substrate in vitro, and possesses a nucleotide-binding pocket capable of binding AMP and cAMP, pointing to a role in cAMP/AMP metabolism or signaling rather than canonical PKA anchoring."},"narrative":{"mechanistic_narrative":"AKAL (PF3D7_0512900) is an atypical AKAP-like protein of Plasmodium falciparum expressed in mature asexual stages, merozoites, and gametocytes [PMID:27129434]. Despite its AKAP-like designation, AKAL does not anchor the PKA regulatory subunit directly; instead it binds Pf14-3-3I, which in turn associates with PfPKA-R, placing AKAL only indirectly within the PKA signaling network [PMID:27129434]. AKAL is itself not a PKA substrate in vitro, whereas its partners Pf14-3-3I and PfPKA-R are phosphorylated by PKA, indicating that regulation of this assembly acts on the partners rather than on AKAL [PMID:27129434]. Consistent with a non-canonical role, AKAL harbors a conserved nucleotide-binding pocket that binds AMP and cAMP, pointing toward a function in cAMP/AMP-related metabolism or signaling rather than classical PKA anchoring [PMID:27129434]. Beyond these interaction and biochemical observations, no functional phenotype or in vivo role for AKAL has been characterized in the available corpus.","teleology":[{"year":2016,"claim":"Establishing where and when AKAL is present was needed before any functional role could be assigned; expression was mapped across parasite life-cycle stages.","evidence":"RT-qPCR and immunofluorescence in P. falciparum 3D7","pmids":["27129434"],"confidence":"Medium","gaps":["No functional consequence tied to the observed stage expression or localization","Subcellular localization not resolved to a specific compartment"],"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"claim":"The central question was whether an AKAP-like protein anchors PKA in P. falciparum; interaction studies showed AKAL does not bind PfPKA-R directly but connects via Pf14-3-3I, redefining it as an atypical, indirect participant in PKA signaling.","evidence":"Reciprocal pull-downs with recombinant AKAL, proteomic identification, and IFA colocalization in P. falciparum 3D7","pmids":["27129434"],"confidence":"Medium","gaps":["Functional significance of the AKAL–Pf14-3-3I interaction in vivo not tested","Stoichiometry and interface of the AKAL–Pf14-3-3I–PfPKA-R assembly unresolved"],"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"claim":"To probe how the complex is regulated, phosphorylation was tested; AKAL is not a PKA substrate whereas Pf14-3-3I and PfPKA-R are, implying control acts on the partners rather than AKAL itself.","evidence":"Radiolabel in vitro kinase assay","pmids":["27129434"],"confidence":"Medium","gaps":["In vitro result not confirmed in cellular context","Whether AKAL is modified by other kinases untested"],"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":2016,"claim":"Whether AKAL has an intrinsic biochemical activity beyond scaffolding was addressed by identifying a nucleotide-binding pocket that binds AMP and cAMP, hinting at a role in cAMP/AMP metabolism or signaling.","evidence":"Qualitative nucleotide-binding experiments supported by a computational structural model","pmids":["27129434"],"confidence":"Low","gaps":["Qualitative binding not validated by quantitative affinity or enzymatic assays","No demonstrated catalytic or signaling output from nucleotide binding","Structural pocket inferred from modeling, not an experimental structure"],"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]},{"year":null,"claim":"The in vivo role of AKAL, the functional consequence of its AMP/cAMP binding, and its contribution to parasite biology remain unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No genetic knockout or knockdown phenotype for AKAL reported","No demonstrated link between nucleotide binding and a metabolic or signaling pathway","No experimental structure of AKAL or its complexes"],"attribution":[{"species":"Plasmodium falciparum","strain":"3D7"}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["AKAL–Pf14-3-3I–PfPKA-R indirect assembly"],"partners":["PF14-3-3I"],"other_free_text":["Atypical AKAP-like protein that does not bind PfPKA-R directly (23% identity to central domain of human AKAP18δ) [#4]","Not a PKA substrate in vitro, unlike its partners Pf14-3-3I and PfPKA-R [#2]","Binds AMP and cAMP via a conserved nucleotide-binding pocket [#3]","Expressed in mature asexual stages, merozoites, and gametocytes [#0]"]}},"prefetch_data":{},"corpus_meta":[{"pmid":"26010880","id":"PMC_26010880","title":"The Stapled AKAP Disruptor Peptide STAD-2 Displays Antimalarial Activity through a PKA-Independent Mechanism.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26010880","citation_count":16,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["AKAP"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"27835919","id":"PMC_27835919","title":"PKA and Apicomplexan Parasite Diseases.","date":"2016","source":"Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme","url":"https://pubmed.ncbi.nlm.nih.gov/27835919","citation_count":9,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["AKAP"],"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":["AKAL","AKAP"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":4680,"output_tokens":1276,"usd":0.01659,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6142,"output_tokens":2154,"usd":0.04228,"stage2_stop_reason":"end_turn"},"total_usd":0.05887,"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\": 2016,\n      \"finding\": \"AKAL (PF3D7_0512900) is expressed in mature asexual stages, merozoites, and gametocytes of P. falciparum 3D7, as determined by RT-qPCR and immunofluorescence assays.\",\n      \"method\": \"RT-qPCR and immunofluorescence assay\",\n      \"journal\": \"Malaria journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — two orthogonal methods (RT-qPCR + IFA) in a single lab, but no functional consequence directly linked to localization\",\n      \"pmids\": [\"27129434\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"AKAL (PF3D7_0512900) does NOT interact directly with the P. falciparum PKA regulatory subunit (PfPKA-R); instead, AKAL binds Pf14-3-3I, which in turn interacts with PfPKA-R, establishing an indirect connection to PKA signaling.\",\n      \"method\": \"Pull-down experiments with recombinant AKAL protein and parasite protein extracts, biochemical and proteomic approaches, immunofluorescence colocalization\",\n      \"journal\": \"Malaria journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal pull-down plus proteomic confirmation and IFA colocalization, single lab\",\n      \"pmids\": [\"27129434\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"AKAL (PF3D7_0512900) is NOT phosphorylated by PKA in vitro, whereas its binding partner Pf14-3-3I and PfPKA-R are phosphorylated by PKA, indicating that phosphorylation of the complex components (but not AKAL itself) may regulate these interactions.\",\n      \"method\": \"Radiolabel in vitro kinase activity assay\",\n      \"journal\": \"Malaria journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — in vitro biochemical assay, single lab, single method\",\n      \"pmids\": [\"27129434\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"AKAL (PF3D7_0512900) binds nucleotides AMP and cAMP, as demonstrated by qualitative nucleotide binding experiments and structural modeling that revealed a conserved nucleotide-binding pocket, suggesting possible involvement in AMPK pathway or phosphodiesterase-associated activities.\",\n      \"method\": \"Structural modeling and qualitative nucleotide binding experiments\",\n      \"journal\": \"Malaria journal\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — qualitative binding assay supported by computational structural model, single lab, no functional validation\",\n      \"pmids\": [\"27129434\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"AKAL (PF3D7_0512900) shares 23% sequence identity with the central domain of human AKAP18δ but represents an atypical AKAP in that it does not anchor PKA-R directly; instead PKA localization in P. falciparum may involve other partners.\",\n      \"method\": \"Sequence alignment and biochemical interaction studies\",\n      \"journal\": \"Malaria journal\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — sequence comparison combined with negative interaction result, single lab\",\n      \"pmids\": [\"27129434\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    }\n  ],\n  \"current_model\": \"AKAL (PF3D7_0512900) is an atypical AKAP-like protein expressed in mature asexual stages, merozoites, and gametocytes of P. falciparum that does not directly bind the PKA regulatory subunit but instead interacts with Pf14-3-3I (which itself binds PfPKA-R), is not a PKA substrate in vitro, and possesses a nucleotide-binding pocket capable of binding AMP and cAMP, pointing to a role in cAMP/AMP metabolism or signaling rather than canonical PKA anchoring.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"AKAL (PF3D7_0512900) is an atypical AKAP-like protein of Plasmodium falciparum expressed in mature asexual stages, merozoites, and gametocytes [#0]. Despite its AKAP-like designation, AKAL does not anchor the PKA regulatory subunit directly; instead it binds Pf14-3-3I, which in turn associates with PfPKA-R, placing AKAL only indirectly within the PKA signaling network [#1]. AKAL is itself not a PKA substrate in vitro, whereas its partners Pf14-3-3I and PfPKA-R are phosphorylated by PKA, indicating that regulation of this assembly acts on the partners rather than on AKAL [#2]. Consistent with a non-canonical role, AKAL harbors a conserved nucleotide-binding pocket that binds AMP and cAMP, pointing toward a function in cAMP/AMP-related metabolism or signaling rather than classical PKA anchoring [#3]. Beyond these interaction and biochemical observations, no functional phenotype or in vivo role for AKAL has been characterized in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 2016,\n      \"claim\": \"Establishing where and when AKAL is present was needed before any functional role could be assigned; expression was mapped across parasite life-cycle stages.\",\n      \"evidence\": \"RT-qPCR and immunofluorescence in P. falciparum 3D7\",\n      \"pmids\": [\"27129434\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No functional consequence tied to the observed stage expression or localization\",\n        \"Subcellular localization not resolved to a specific compartment\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"The central question was whether an AKAP-like protein anchors PKA in P. falciparum; interaction studies showed AKAL does not bind PfPKA-R directly but connects via Pf14-3-3I, redefining it as an atypical, indirect participant in PKA signaling.\",\n      \"evidence\": \"Reciprocal pull-downs with recombinant AKAL, proteomic identification, and IFA colocalization in P. falciparum 3D7\",\n      \"pmids\": [\"27129434\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Functional significance of the AKAL\\u2013Pf14-3-3I interaction in vivo not tested\",\n        \"Stoichiometry and interface of the AKAL\\u2013Pf14-3-3I\\u2013PfPKA-R assembly unresolved\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"To probe how the complex is regulated, phosphorylation was tested; AKAL is not a PKA substrate whereas Pf14-3-3I and PfPKA-R are, implying control acts on the partners rather than AKAL itself.\",\n      \"evidence\": \"Radiolabel in vitro kinase assay\",\n      \"pmids\": [\"27129434\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"In vitro result not confirmed in cellular context\",\n        \"Whether AKAL is modified by other kinases untested\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Whether AKAL has an intrinsic biochemical activity beyond scaffolding was addressed by identifying a nucleotide-binding pocket that binds AMP and cAMP, hinting at a role in cAMP/AMP metabolism or signaling.\",\n      \"evidence\": \"Qualitative nucleotide-binding experiments supported by a computational structural model\",\n      \"pmids\": [\"27129434\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Qualitative binding not validated by quantitative affinity or enzymatic assays\",\n        \"No demonstrated catalytic or signaling output from nucleotide binding\",\n        \"Structural pocket inferred from modeling, not an experimental structure\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The in vivo role of AKAL, the functional consequence of its AMP/cAMP binding, and its contribution to parasite biology remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No genetic knockout or knockdown phenotype for AKAL reported\",\n        \"No demonstrated link between nucleotide binding and a metabolic or signaling pathway\",\n        \"No experimental structure of AKAL or its complexes\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"3D7\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0000287\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0030552\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005515\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"GO:0007165\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"complexes\": [\n      \"AKAL\\u2013Pf14-3-3I\\u2013PfPKA-R indirect assembly\"\n    ],\n    \"partners\": [\n      \"Pf14-3-3I\"\n    ],\n    \"other_free_text\": [\n      \"Atypical AKAP-like protein that does not bind PfPKA-R directly (23% identity to central domain of human AKAP18\\u03b4) [#4]\",\n      \"Not a PKA substrate in vitro, unlike its partners Pf14-3-3I and PfPKA-R [#2]\",\n      \"Binds AMP and cAMP via a conserved nucleotide-binding pocket [#3]\",\n      \"Expressed in mature asexual stages, merozoites, and gametocytes [#0]\"\n    ]\n  }\n}","audit_flag":null,"strains":[{"strain":"Pf3D7","gene_id":"PF3D7_0512900","name":null,"product":"AKAP-like protein","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PbANKA","gene_id":"PBANKA_1112600","name":null,"product":"AKAP-like protein, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PkH","gene_id":"PKNH_1020200","name":null,"product":"AKAP-like protein, putative","uniprot_acc":null,"chromosome":null,"start_pos":null}],"gene_symbol":"AKAL","evaluation":null}