{"gene":"PF3D7_1330600","run_date":"2026-09-08T14:19:06","timeline":{"discoveries":[{"year":2005,"finding":"The tufA gene encoded by the P. falciparum apicoplast genome is translationally active; its protein product EF-Tu was detected and localized within the apicoplast organelle, with maximal expression in the trophozoite stage of the intraerythrocytic cycle.","method":"Protein localization (immunofluorescence/fractionation), stage-specific expression analysis","journal":"Molecular microbiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization experiment with organelle fractionation and stage-specific expression, single lab with multiple methods","pmids":["15773980"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2005,"finding":"The drug thiostrepton, which binds the apicoplast LSU rRNA, reduced P. falciparum apicoplast EF-Tu (tufA) protein levels, providing evidence that apicoplast translation is the site of action of this drug.","method":"Drug treatment followed by western blot quantification of EF-Tu protein levels","journal":"Molecular microbiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct biochemical measurement of protein levels under drug treatment, single lab","pmids":["15773980"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":1999,"finding":"Recombinant P. falciparum apicoplast EF-Tu (EF-Tu(pl)) forms demonstrable complexes in vitro with the EF-Tu inhibitors kirromycin, GE2270A, and enacyloxin IIa, which bind to different sites on EF-Tu; these inhibitors show antimalarial activity in blood cultures.","method":"In vitro complex formation assay with recombinant protein and structurally distinct antibiotic inhibitors; blood culture antimalarial assay","journal":"Protist","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with recombinant protein and multiple inhibitor classes, single lab","pmids":["10505418"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2010,"finding":"Recombinant P. falciparum apicoplast EF-Tu (PfEF-Tu, encoded by tufA) hydrolyses GTP and interacts with the nuclear-encoded elongation factor PfEF-Ts; PfEF-Ts mediates nucleotide exchange on PfEF-Tu (GDP release), as shown by in vitro assays and homology modeling of the PfEF-Tu:PfEF-Ts complex.","method":"Recombinant protein production, GTPase assay, in vitro nucleotide exchange assay, Co-IP/pulldown interaction assay, homology modeling","journal":"International journal for parasitology","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstituted enzymatic activity (GTPase), direct interaction assay, nucleotide exchange assay, and structural modeling all in one study with multiple orthogonal methods","pmids":["21163263"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2010,"finding":"The EF-Tu inhibitor kirromycin affects PfEF-Tu GTPase/translation activity in vitro, confirming apicoplast EF-Tu as the molecular target of this drug in P. falciparum.","method":"In vitro enzymatic inhibition assay with recombinant PfEF-Tu and kirromycin","journal":"International journal for parasitology","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with recombinant protein and drug, single lab","pmids":["21163263"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2010,"finding":"P. falciparum apicoplast EF-Tu (PfEF-Tu) possesses chaperone-related disulphide reductase activity, providing a functional rationale for retention of the tufA gene on the plastid genome.","method":"In vitro biochemical assay for disulphide reductase activity using recombinant PfEF-Tu","journal":"International journal for parasitology","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — in vitro enzymatic assay with recombinant protein, novel activity, single lab","pmids":["21163263"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"current_model":"PF3D7_1330600 encodes the apicoplast-encoded elongation factor Tu (EF-Tu/tufA) in P. falciparum, which is translationally active within the apicoplast (maximal in trophozoites), hydrolyses GTP, forms a functional complex with the nuclear-encoded EF-Ts (which mediates GDP release/nucleotide exchange on EF-Tu), possesses disulphide reductase activity, and is directly targeted by antibiotic inhibitors including kirromycin, GE2270A, enacyloxin IIa, and thiostrepton."},"narrative":{"mechanistic_narrative":"PF3D7_1330600 (tufA) encodes the apicoplast-encoded elongation factor Tu (EF-Tu) of Plasmodium falciparum, a translationally active component of the organellar protein synthesis machinery whose product is localized within the apicoplast and maximally expressed in the trophozoite stage [PMID:15773980]. As a canonical translation GTPase, recombinant PfEF-Tu hydrolyses GTP and undergoes nucleotide exchange (GDP release) mediated by the nuclear-encoded PfEF-Ts, with which it forms a functional complex [PMID:21163263]. Beyond its translational role, PfEF-Tu displays an additional chaperone-related disulphide reductase activity, offering a rationale for retention of tufA on the plastid genome [PMID:21163263]. The protein is a validated antimalarial target: it is directly bound and inhibited by structurally distinct EF-Tu antibiotics kirromycin, GE2270A, and enacyloxin IIa in vitro [PMID:10505418, PMID:21163263], and its levels are depleted by thiostrepton, which targets apicoplast translation [PMID:15773980].","teleology":[{"year":1999,"claim":"The first question was whether the apicoplast EF-Tu is a genuine drug target — reconstitution with recombinant protein showed it directly engages multiple classes of EF-Tu antibiotics that also kill parasites, establishing apicoplast translation as an antimalarial vulnerability.","evidence":"In vitro complex-formation assays with recombinant EF-Tu(pl) and kirromycin, GE2270A, enacyloxin IIa, plus blood-culture antimalarial assays","pmids":["10505418"],"confidence":"Medium","gaps":["In vitro binding does not demonstrate that drug killing in culture proceeds through EF-Tu specifically","No enzymatic readout of inhibition at this stage","No structural mapping of the distinct binding sites"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2005,"claim":"It was unknown whether the plastid tufA gene is actually expressed as protein and where — localization and stage-specific analysis showed EF-Tu is produced and resides in the apicoplast with peak trophozoite expression, confirming an active organellar translation apparatus.","evidence":"Protein localization by immunofluorescence/fractionation and stage-specific expression analysis in P. falciparum","pmids":["15773980"],"confidence":"Medium","gaps":["Does not define the full set of translation partners in the organelle","Stage-specific regulation mechanism not addressed"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2005,"claim":"To link a known antibiotic to apicoplast translation, thiostrepton treatment was shown to reduce EF-Tu protein levels, providing cellular evidence that this drug acts at the site of apicoplast protein synthesis.","evidence":"Drug treatment followed by western-blot quantification of EF-Tu protein levels in P. falciparum","pmids":["15773980"],"confidence":"Medium","gaps":["Protein-level reduction does not distinguish direct from downstream effects on EF-Tu","Thiostrepton targets LSU rRNA, not EF-Tu itself"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2010,"claim":"The catalytic and partner biochemistry was resolved — reconstituted PfEF-Tu hydrolyses GTP, physically interacts with nuclear-encoded PfEF-Ts, and undergoes PfEF-Ts-mediated GDP release, establishing a functional EF-Tu/EF-Ts nucleotide-exchange cycle across the two genomes.","evidence":"Recombinant GTPase, nucleotide-exchange, and Co-IP/pulldown assays plus homology modeling of the PfEF-Tu:PfEF-Ts complex","pmids":["21163263"],"confidence":"High","gaps":["Interaction with the ternary complex (aminoacyl-tRNA) not directly demonstrated","Complex geometry rests on homology modeling, not an experimental structure"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2010,"claim":"The antibiotic-target link was closed at the enzymatic level — kirromycin was shown to inhibit PfEF-Tu GTPase/translation activity in vitro, confirming EF-Tu as the direct molecular target rather than a bystander.","evidence":"In vitro enzymatic inhibition assay with recombinant PfEF-Tu and kirromycin","pmids":["21163263"],"confidence":"Medium","gaps":["Enzymatic inhibition not yet tied to a defined resistance mutation in the parasite","Only kirromycin tested enzymatically among the inhibitor panel"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2010,"claim":"A moonlighting activity was uncovered — recombinant PfEF-Tu exhibits chaperone-related disulphide reductase activity, offering a functional explanation for why tufA is retained on the plastid genome.","evidence":"In vitro disulphide reductase biochemical assay with recombinant PfEF-Tu","pmids":["21163263"],"confidence":"Medium","gaps":["Physiological substrates of the reductase activity not identified","In vivo relevance of this activity not tested"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":null,"claim":"Whether PfEF-Tu is genetically essential, how its stage-regulated expression is controlled, and the in vivo role and substrates of its disulphide reductase activity remain unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No genetic knockout/knockdown of tufA in the corpus","No experimental structure of PfEF-Tu or its complexes","Resistance-conferring mutations to the antibiotic inhibitors not mapped"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["PfEF-Tu:PfEF-Ts complex (with nuclear-encoded elongation factor Ts) [#3]"],"partners":["PFEF-TS (NUCLEAR-ENCODED ELONGATION FACTOR TS; NUCLEOTIDE-EXCHANGE FACTOR) [#3]","KIRROMYCIN (INHIBITOR) [#2, #4]","GE2270A (INHIBITOR) [#2]","ENACYLOXIN IIA (INHIBITOR) [#2]"],"other_free_text":["Apicoplast-encoded (tufA) elongation factor Tu; expression maximal in the trophozoite stage [#0]","Thiostrepton (apicoplast LSU rRNA-binding drug) reduces EF-Tu protein levels, marking apicoplast translation as its site of action [#1]","Validated antimalarial target inhibited by structurally distinct EF-Tu antibiotics that bind different sites [#2, #4]","Chaperone-related disulphide reductase activity proposed as rationale for retention of tufA on the plastid genome [#5]"]}},"prefetch_data":{},"corpus_meta":[{"pmid":"15684081","id":"PMC_15684081","title":"A monkey's tale: the origin of Plasmodium vivax as a human malaria parasite.","date":"2005","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/15684081","citation_count":158,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["elongation factor Tu"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"21741312","id":"PMC_21741312","title":"Protein translation in Plasmodium parasites.","date":"2011","source":"Trends in parasitology","url":"https://pubmed.ncbi.nlm.nih.gov/21741312","citation_count":74,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["curated_ref"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"15773980","id":"PMC_15773980","title":"The apicoplast of Plasmodium falciparum is translationally active.","date":"2005","source":"Molecular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/15773980","citation_count":59,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["elongation factor Tu"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"10505418","id":"PMC_10505418","title":"Antibiotic inhibitors of organellar protein synthesis in Plasmodium falciparum.","date":"1999","source":"Protist","url":"https://pubmed.ncbi.nlm.nih.gov/10505418","citation_count":56,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["elongation factor Tu"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"19818838","id":"PMC_19818838","title":"Phylogeny of Asian primate malaria parasites inferred from apicoplast genome-encoded genes with special emphasis on the positions of Plasmodium vivax and P. fragile.","date":"2010","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/19818838","citation_count":17,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["elongation factor Tu"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"21163263","id":"PMC_21163263","title":"Interaction of apicoplast-encoded elongation factor (EF) EF-Tu with nuclear-encoded EF-Ts mediates translation in the Plasmodiumfalciparum plastid.","date":"2010","source":"International journal for parasitology","url":"https://pubmed.ncbi.nlm.nih.gov/21163263","citation_count":17,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["curated_ref"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"17602895","id":"PMC_17602895","title":"Analysis of elongation factor Tu (tuf A) of apicoplast from Indian Plasmodium vivax isolates.","date":"2007","source":"Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases","url":"https://pubmed.ncbi.nlm.nih.gov/17602895","citation_count":8,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["elongation factor Tu"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":5909,"output_tokens":1570,"usd":0.020639,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6519,"output_tokens":3157,"usd":0.05576,"stage2_stop_reason":"end_turn"},"total_usd":0.076399,"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\": 2005,\n      \"finding\": \"The tufA gene encoded by the P. falciparum apicoplast genome is translationally active; its protein product EF-Tu was detected and localized within the apicoplast organelle, with maximal expression in the trophozoite stage of the intraerythrocytic cycle.\",\n      \"method\": \"Protein localization (immunofluorescence/fractionation), stage-specific expression analysis\",\n      \"journal\": \"Molecular microbiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiment with organelle fractionation and stage-specific expression, single lab with multiple methods\",\n      \"pmids\": [\"15773980\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"The drug thiostrepton, which binds the apicoplast LSU rRNA, reduced P. falciparum apicoplast EF-Tu (tufA) protein levels, providing evidence that apicoplast translation is the site of action of this drug.\",\n      \"method\": \"Drug treatment followed by western blot quantification of EF-Tu protein levels\",\n      \"journal\": \"Molecular microbiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct biochemical measurement of protein levels under drug treatment, single lab\",\n      \"pmids\": [\"15773980\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Recombinant P. falciparum apicoplast EF-Tu (EF-Tu(pl)) forms demonstrable complexes in vitro with the EF-Tu inhibitors kirromycin, GE2270A, and enacyloxin IIa, which bind to different sites on EF-Tu; these inhibitors show antimalarial activity in blood cultures.\",\n      \"method\": \"In vitro complex formation assay with recombinant protein and structurally distinct antibiotic inhibitors; blood culture antimalarial assay\",\n      \"journal\": \"Protist\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with recombinant protein and multiple inhibitor classes, single lab\",\n      \"pmids\": [\"10505418\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Recombinant P. falciparum apicoplast EF-Tu (PfEF-Tu, encoded by tufA) hydrolyses GTP and interacts with the nuclear-encoded elongation factor PfEF-Ts; PfEF-Ts mediates nucleotide exchange on PfEF-Tu (GDP release), as shown by in vitro assays and homology modeling of the PfEF-Tu:PfEF-Ts complex.\",\n      \"method\": \"Recombinant protein production, GTPase assay, in vitro nucleotide exchange assay, Co-IP/pulldown interaction assay, homology modeling\",\n      \"journal\": \"International journal for parasitology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstituted enzymatic activity (GTPase), direct interaction assay, nucleotide exchange assay, and structural modeling all in one study with multiple orthogonal methods\",\n      \"pmids\": [\"21163263\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"The EF-Tu inhibitor kirromycin affects PfEF-Tu GTPase/translation activity in vitro, confirming apicoplast EF-Tu as the molecular target of this drug in P. falciparum.\",\n      \"method\": \"In vitro enzymatic inhibition assay with recombinant PfEF-Tu and kirromycin\",\n      \"journal\": \"International journal for parasitology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with recombinant protein and drug, single lab\",\n      \"pmids\": [\"21163263\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"P. falciparum apicoplast EF-Tu (PfEF-Tu) possesses chaperone-related disulphide reductase activity, providing a functional rationale for retention of the tufA gene on the plastid genome.\",\n      \"method\": \"In vitro biochemical assay for disulphide reductase activity using recombinant PfEF-Tu\",\n      \"journal\": \"International journal for parasitology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro enzymatic assay with recombinant protein, novel activity, single lab\",\n      \"pmids\": [\"21163263\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    }\n  ],\n  \"current_model\": \"PF3D7_1330600 encodes the apicoplast-encoded elongation factor Tu (EF-Tu/tufA) in P. falciparum, which is translationally active within the apicoplast (maximal in trophozoites), hydrolyses GTP, forms a functional complex with the nuclear-encoded EF-Ts (which mediates GDP release/nucleotide exchange on EF-Tu), possesses disulphide reductase activity, and is directly targeted by antibiotic inhibitors including kirromycin, GE2270A, enacyloxin IIa, and thiostrepton.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"PF3D7_1330600 (tufA) encodes the apicoplast-encoded elongation factor Tu (EF-Tu) of Plasmodium falciparum, a translationally active component of the organellar protein synthesis machinery whose product is localized within the apicoplast and maximally expressed in the trophozoite stage [#0]. As a canonical translation GTPase, recombinant PfEF-Tu hydrolyses GTP and undergoes nucleotide exchange (GDP release) mediated by the nuclear-encoded PfEF-Ts, with which it forms a functional complex [#3]. Beyond its translational role, PfEF-Tu displays an additional chaperone-related disulphide reductase activity, offering a rationale for retention of tufA on the plastid genome [#5]. The protein is a validated antimalarial target: it is directly bound and inhibited by structurally distinct EF-Tu antibiotics kirromycin, GE2270A, and enacyloxin IIa in vitro [#2, #4], and its levels are depleted by thiostrepton, which targets apicoplast translation [#1].\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"The first question was whether the apicoplast EF-Tu is a genuine drug target — reconstitution with recombinant protein showed it directly engages multiple classes of EF-Tu antibiotics that also kill parasites, establishing apicoplast translation as an antimalarial vulnerability.\",\n      \"evidence\": \"In vitro complex-formation assays with recombinant EF-Tu(pl) and kirromycin, GE2270A, enacyloxin IIa, plus blood-culture antimalarial assays\",\n      \"pmids\": [\"10505418\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\n        \"In vitro binding does not demonstrate that drug killing in culture proceeds through EF-Tu specifically\",\n        \"No enzymatic readout of inhibition at this stage\",\n        \"No structural mapping of the distinct binding sites\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"It was unknown whether the plastid tufA gene is actually expressed as protein and where — localization and stage-specific analysis showed EF-Tu is produced and resides in the apicoplast with peak trophozoite expression, confirming an active organellar translation apparatus.\",\n      \"evidence\": \"Protein localization by immunofluorescence/fractionation and stage-specific expression analysis in P. falciparum\",\n      \"pmids\": [\"15773980\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\n        \"Does not define the full set of translation partners in the organelle\",\n        \"Stage-specific regulation mechanism not addressed\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"To link a known antibiotic to apicoplast translation, thiostrepton treatment was shown to reduce EF-Tu protein levels, providing cellular evidence that this drug acts at the site of apicoplast protein synthesis.\",\n      \"evidence\": \"Drug treatment followed by western-blot quantification of EF-Tu protein levels in P. falciparum\",\n      \"pmids\": [\"15773980\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\n        \"Protein-level reduction does not distinguish direct from downstream effects on EF-Tu\",\n        \"Thiostrepton targets LSU rRNA, not EF-Tu itself\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"The catalytic and partner biochemistry was resolved — reconstituted PfEF-Tu hydrolyses GTP, physically interacts with nuclear-encoded PfEF-Ts, and undergoes PfEF-Ts-mediated GDP release, establishing a functional EF-Tu/EF-Ts nucleotide-exchange cycle across the two genomes.\",\n      \"evidence\": \"Recombinant GTPase, nucleotide-exchange, and Co-IP/pulldown assays plus homology modeling of the PfEF-Tu:PfEF-Ts complex\",\n      \"pmids\": [\"21163263\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\n        \"Interaction with the ternary complex (aminoacyl-tRNA) not directly demonstrated\",\n        \"Complex geometry rests on homology modeling, not an experimental structure\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"The antibiotic-target link was closed at the enzymatic level — kirromycin was shown to inhibit PfEF-Tu GTPase/translation activity in vitro, confirming EF-Tu as the direct molecular target rather than a bystander.\",\n      \"evidence\": \"In vitro enzymatic inhibition assay with recombinant PfEF-Tu and kirromycin\",\n      \"pmids\": [\"21163263\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\n        \"Enzymatic inhibition not yet tied to a defined resistance mutation in the parasite\",\n        \"Only kirromycin tested enzymatically among the inhibitor panel\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"A moonlighting activity was uncovered — recombinant PfEF-Tu exhibits chaperone-related disulphide reductase activity, offering a functional explanation for why tufA is retained on the plastid genome.\",\n      \"evidence\": \"In vitro disulphide reductase biochemical assay with recombinant PfEF-Tu\",\n      \"pmids\": [\"21163263\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\n        \"Physiological substrates of the reductase activity not identified\",\n        \"In vivo relevance of this activity not tested\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Whether PfEF-Tu is genetically essential, how its stage-regulated expression is controlled, and the in vivo role and substrates of its disulphide reductase activity remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\n        \"No genetic knockout/knockdown of tufA in the corpus\",\n        \"No experimental structure of PfEF-Tu or its complexes\",\n        \"Resistance-conferring mutations to the antibiotic inhibitors not mapped\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003924\", \"supporting_discovery_ids\": [3, 4]},\n      {\"term_id\": \"GO:0003746\", \"supporting_discovery_ids\": [0, 3]},\n      {\"term_id\": \"GO:0005525\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0015036\", \"supporting_discovery_ids\": [5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0009507\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0020011\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0006414\", \"supporting_discovery_ids\": [0, 1, 3]}\n    ],\n    \"complexes\": [\n      \"PfEF-Tu:PfEF-Ts complex (with nuclear-encoded elongation factor Ts) [#3]\"\n    ],\n    \"partners\": [\n      \"PfEF-Ts (nuclear-encoded elongation factor Ts; nucleotide-exchange factor) [#3]\",\n      \"kirromycin (inhibitor) [#2, #4]\",\n      \"GE2270A (inhibitor) [#2]\",\n      \"enacyloxin IIa (inhibitor) [#2]\"\n    ],\n    \"other_free_text\": [\n      \"Apicoplast-encoded (tufA) elongation factor Tu; expression maximal in the trophozoite stage [#0]\",\n      \"Thiostrepton (apicoplast LSU rRNA-binding drug) reduces EF-Tu protein levels, marking apicoplast translation as its site of action [#1]\",\n      \"Validated antimalarial target inhibited by structurally distinct EF-Tu antibiotics that bind different sites [#2, #4]\",\n      \"Chaperone-related disulphide reductase activity proposed as rationale for retention of tufA on the plastid genome [#5]\"\n    ]\n  }\n}","audit_flag":null,"strains":[{"strain":"Pf3D7","gene_id":"PF3D7_1330600","name":null,"product":"elongation factor Tu, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PbANKA","gene_id":"PBANKA_1345600","name":null,"product":"elongation factor Tu, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PkH","gene_id":"PKNH_1269800","name":null,"product":"elongation factor Tu, putative","uniprot_acc":null,"chromosome":null,"start_pos":null}],"gene_symbol":"","evaluation":null}