{"gene":"PF3D7_1027700","run_date":"2026-09-08T16:26:55","timeline":{"discoveries":[{"year":2008,"finding":"CEN3 (PF3D7_1027700) protein is expressed in sporozoites, asexual blood forms, gametocytes, and oocysts, and localizes in close proximity to the nucleus of sporozoites and asexual schizonts, with immunofluorescence double-staining with alpha-tubulin indicating association with the parasite centrosome (centriolar plaque).","method":"Immunoelectron microscopy and confocal/widefield immunofluorescence microscopy with anti-centrin and anti-alpha-tubulin antibodies in P. falciparum","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct localization by immunofluorescence and immunoelectron microscopy in a single study, but antibody used recognizes multiple centrins and does not exclusively report on CEN3","pmids":["18693242"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2021,"finding":"CEN3 (PF3D7_1027700) localizes to the extranuclear compartment of the centriolar plaque in P. falciparum blood-stage schizonts, as established using a polyclonal rabbit antibody raised against recombinant PfCen3-6His (PF3D7_1027700.1); this extranuclear position is distinct from the intranuclear compartment where microtubules are nucleated.","method":"Super-resolution STED microscopy and correlative light/electron tomography with affinity-purified anti-PfCentrin3 antibody in P. falciparum NF54 strain","journal":"Life science alliance","confidence":"High","confidence_rationale":"Tier 2 / Moderate — antibody validated by western blot against recombinant PfCen3-6His, localization confirmed by super-resolution STED and correlative electron tomography, single lab but multiple orthogonal imaging methods","pmids":["34535568"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":"NF54"}]},{"year":2023,"finding":"CEN3 (PF3D7_1027700) undergoes calcium-inducible liquid-liquid phase separation (LLPS) in vitro: recombinant PfCen3 forms droplets upon Ca2+ addition that display fusion and surface wetting; unlike PfCen1, PfCen3 condensates show partial irreversibility (maturation towards a gel-like state) upon EDTA addition, a process slowed at lower temperature. Mg2+ causes only a reduced response compared to Ca2+, indicating calcium-specificity. The disordered N-terminus and calcium-binding EF-hand motifs are defined as essential features for this condensation.","method":"In vitro recombinant protein phase separation assay (6His-tagged and untagged PfCen3 expressed in E. coli), turbidity/light-scattering measurements, live droplet imaging including fusion-event analysis at varying temperatures, EDTA dissolution assays","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with multiple orthogonal quantitative readouts (turbidity, imaging, temperature variation, tagged and untagged protein), single lab but rigorous biochemical controls including calcium specificity and concentration-dependence","pmids":["38150475"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2023,"finding":"CEN3 (PF3D7_1027700) was identified as a specific co-immunoprecipitation hit with PfCen1-GFP in P. falciparum, indicating a physical interaction between CEN3 and CEN1 at the centriolar plaque; CEN3 co-localizes with the Sfi1-like centriolar plaque protein PfSlp at the extranuclear centriolar plaque as shown by dual-color STED microscopy.","method":"Co-immunoprecipitation (PfCen1-GFP pulldown with anti-PfCen3 as readout), reciprocal IP, dual-color STED super-resolution microscopy with anti-PfCen3 antibody in P. falciparum","journal":"PLoS pathogens","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — reciprocal Co-IP and super-resolution imaging in single lab, but no cross-linking and quantitative enrichment analysis not performed; interaction corroborated by previously published yeast two-hybrid data cited in abstract","pmids":["37130129"],"is_preprint":false,"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2019,"finding":"In P. berghei, deletion of cen-3 (ortholog of PF3D7_1027700) could not be achieved, indicating that CEN3 is likely essential for asexual blood-stage development. Additionally, CEN-3 forms part of a multi-protein complex with other centrins (CEN-1, CEN-2, CEN-4) as shown by complex formation experiments with PbCEN-4.","method":"Gene deletion attempts in P. berghei (failed knockout indicating essentiality); multi-protein complex formation analysis with other centrins","journal":"Biology open","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — essentiality inferred from failed deletion attempts (single lab), complex formation reported without full mechanistic detail in abstract; consistent with inability to delete cen-1 and cen-2 as well","pmids":["30541825"],"is_preprint":false,"attribution":[{"species":"Plasmodium berghei","strain":""}]}],"current_model":"CEN3 (PF3D7_1027700) is an essential calcium-binding centrin that localizes to the extranuclear compartment of the P. falciparum centriolar plaque, physically interacts with CEN1 and other centrins in a multi-protein complex, and undergoes calcium-induced liquid-liquid phase separation in vitro with partial irreversible condensate maturation, suggesting that Ca2+-regulated biomolecular condensation drives its accumulation at the centrosome during cell division."},"narrative":{"mechanistic_narrative":"CEN3 (PF3D7_1027700) is a calcium-binding centrin that associates with the centrosome-equivalent centriolar plaque of Plasmodium during cell division [PMID:18693242, PMID:34535568]. It is expressed across sporozoite, asexual blood, gametocyte, and oocyst stages and localizes in close proximity to the nucleus, specifically to the extranuclear compartment of the centriolar plaque — a position distinct from the intranuclear region where microtubules are nucleated [PMID:18693242, PMID:34535568]. At this site CEN3 physically interacts with CEN1 (by reciprocal co-immunoprecipitation) and co-localizes with the Sfi1-like plaque protein PfSlp, and in the P. berghei ortholog it forms part of a multi-protein centrin complex including CEN-1, CEN-2, and CEN-4 [PMID:37130129, PMID:30541825]. In P. berghei, cen-3 could not be deleted, indicating the gene is essential for asexual blood-stage development [PMID:30541825]. Biochemically, recombinant CEN3 undergoes calcium-inducible liquid-liquid phase separation in vitro, forming droplets that fuse and wet surfaces; unlike CEN1, CEN3 condensates mature partially irreversibly toward a gel-like state, a temperature-dependent process, and this behavior depends on its disordered N-terminus and EF-hand calcium-binding motifs, establishing Ca2+-specific condensation as a defining property [PMID:38150475]. Beyond its centriolar-plaque localization, essentiality, centrin-complex membership, and calcium-driven condensation, the precise cellular function of CEN3 in centriolar plaque biology has not been mechanistically resolved in the available corpus.","teleology":[{"year":2008,"claim":"Established that CEN3 is a broadly expressed centrin positioned at the parasite centrosome, placing it at the machinery of cell division.","evidence":"Immunoelectron and confocal/widefield immunofluorescence microscopy with anti-centrin and anti-alpha-tubulin antibodies in P. falciparum","pmids":["18693242"],"confidence":"Medium","gaps":["Antibody recognizes multiple centrins and does not exclusively report CEN3","No functional role assigned beyond spatial association with the centrosome","Sub-compartment within the centriolar plaque not resolved"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2019,"claim":"Tested whether CEN3 is dispensable and whether it acts alone; the ortholog proved essential and was shown to sit within a multi-centrin complex.","evidence":"Failed cen-3 gene deletion (essentiality inference) and multi-protein complex formation analysis with PbCEN-4 in P. berghei","pmids":["30541825"],"confidence":"Medium","gaps":["Essentiality inferred from failed knockout rather than conditional depletion","Stoichiometry and architecture of the centrin complex not defined","Direct versus indirect nature of CEN3 contacts within the complex unresolved"],"attribution":[{"species":"Plasmodium berghei","strain":""}]},{"year":2021,"claim":"Resolved where within the centriolar plaque CEN3 acts, distinguishing its extranuclear position from the intranuclear microtubule-nucleating compartment.","evidence":"Super-resolution STED microscopy and correlative light/electron tomography with affinity-purified anti-PfCentrin3 antibody in P. falciparum NF54","pmids":["34535568"],"confidence":"High","gaps":["Functional consequence of extranuclear localization not established","Dynamics of CEN3 recruitment during the cell cycle not tracked","Molecular tethers holding CEN3 in the extranuclear compartment unknown"],"attribution":[{"species":"Plasmodium falciparum","strain":"NF54"}]},{"year":2023,"claim":"Defined CEN3's physical partners at the plaque, connecting it to CEN1 and the Sfi1-like protein PfSlp.","evidence":"Reciprocal co-immunoprecipitation (PfCen1-GFP pulldown) and dual-color STED super-resolution microscopy in P. falciparum","pmids":["37130129"],"confidence":"Medium","gaps":["No cross-linking or quantitative enrichment analysis performed","Whether the CEN3-CEN1 interaction is direct not shown","Functional role of the CEN3-PfSlp co-localization not tested"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":2023,"claim":"Provided a biophysical mechanism for CEN3 accumulation by showing calcium-specific, EF-hand- and N-terminus-dependent phase separation, and distinguished CEN3 from CEN1 by its partial condensate irreversibility.","evidence":"In vitro reconstitution with recombinant tagged and untagged PfCen3, turbidity/light-scattering, live droplet fusion imaging at varying temperatures, and EDTA dissolution assays","pmids":["38150475"],"confidence":"High","gaps":["Phase separation demonstrated in vitro only, not confirmed at the centriolar plaque in vivo","Physiological consequence of irreversible condensate maturation unknown","Whether CEN3 condensation recruits or scaffolds its complex partners not tested"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]},{"year":null,"claim":"How Ca2+-driven CEN3 condensation is coupled to its essential role in centriolar plaque function and parasite division remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No conditional depletion linking loss of CEN3 to a specific division defect","In vitro phase separation not yet connected to in vivo condensate formation","Direct molecular activity of CEN3 within the plaque undefined"],"attribution":[{"species":"Plasmodium falciparum","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["Centrin multi-protein complex (CEN-1, CEN-2, CEN-3, CEN-4) at the centriolar plaque"],"partners":["CEN1 (PFCEN1)","PFSLP (SFI1-LIKE CENTRIOLAR PLAQUE PROTEIN)","PBCEN-1, PBCEN-2, PBCEN-4 (IN P. BERGHEI)"],"other_free_text":["Undergoes calcium-inducible liquid-liquid phase separation in vitro; condensates mature partially irreversibly toward a gel-like state, distinguishing CEN3 from CEN1","Condensation depends on the disordered N-terminus and EF-hand calcium-binding motifs; calcium-specific (Mg2+ gives reduced response)","Localizes to the extranuclear compartment of the centriolar plaque, distinct from the intranuclear microtubule-nucleating region","Essential in P. berghei asexual blood stage (cen-3 deletion could not be achieved)","Expressed in sporozoites, asexual blood forms, gametocytes, and oocysts"]}},"prefetch_data":{},"corpus_meta":[{"pmid":"18693242","id":"PMC_18693242","title":"Centrins, cell cycle regulation proteins in human malaria parasite Plasmodium falciparum.","date":"2008","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/18693242","citation_count":68,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["curated_ref"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"38150475","id":"","title":"Malaria parasite centrins can assemble by Ca2+-inducible condensation.","date":"2023","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/38150475","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["CEN3"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"37130129","id":"","title":"An Sfi1-like centrin-interacting centriolar plaque protein affects nuclear microtubule homeostasis.","date":"2023","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/37130129","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["CEN3"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"34535568","id":"","title":"An extended DNA-free intranuclear compartment organizes centrosome microtubules in malaria parasites.","date":"2021","source":"Life science alliance","url":"https://pubmed.ncbi.nlm.nih.gov/34535568","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["CEN3"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"30541825","id":"","title":"Plasmodium centrin PbCEN-4 localizes to the putative MTOC and is dispensable for malaria parasite proliferation.","date":"2019","source":"Biology open","url":"https://pubmed.ncbi.nlm.nih.gov/30541825","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["CEN3"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"22912579","id":"","title":"Arrest of nuclear division in Plasmodium through blockage of erythrocyte surface exposed ribosomal protein P2.","date":"2012","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/22912579","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["CEN3"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false},{"pmid":"20227667","id":"","title":"Functional identification of the Plasmodium centromere and generation of a Plasmodium artificial chromosome.","date":"2010","source":"Cell host & microbe","url":"https://pubmed.ncbi.nlm.nih.gov/20227667","citation_count":0,"is_preprint":false,"source_species":["Pf"],"role":"primary","match_terms":["CEN3"],"competing_organism_flag":false,"competing_organism_match":"","competing_organism_only":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":9678,"output_tokens":2298,"usd":0.031752,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":6789,"output_tokens":2869,"usd":0.052835,"stage2_stop_reason":"end_turn"},"total_usd":0.084587,"stage1_batch_id":"msgbatch_01NE1MTa8x1hW7Lo4tAaNzY1","stage2_batch_id":"msgbatch_0141vaQdeFvbo2q39AinvXM9","note":"batch pricing = 50% of standard"},"stage1_raw":"I need to carefully analyze each paper to determine what was experimentally established about PF3D7_1027700 (CEN3/centrin-3) specifically.\n\n**Paper [1] PMID:18693242**: Studies all four PfCENs. Immunofluorescence and immunoelectron microscopy show centrin associates with parasite centrosome/nucleus vicinity. Growth complementation in Leishmania centrin-KO — but which PfCEN restored growth is not specified as CEN3. The paper characterizes all four together. I need to be careful not to attribute findings about \"PfCENs\" generically to CEN3 specifically.\n\n**Paper [2] PMID:38150475**: Body text explicitly states PfCen3 undergoes Ca2+-inducible LLPS in vitro (recombinant protein), with partially irreversible condensation (maturation). Disordered N-terminus and Ca2+-binding motifs are essential features.\n\n**Paper [3] PMID:37130129**: PfCen3 antibody used to label the extranuclear centriolar plaque. Co-IP of PfCen1-GFP identified PfCen3 as a specific hit. CEN3 localizes to extranuclear centriolar plaque.\n\n**Paper [4] PMID:34535568**: Anti-PfCentrin3 antibody generated and used; centrin is extranuclear at centriolar plaque. This confirms CEN3 localization experimentally.\n\n**Paper [5] PMID:30541825**: Attempts to delete cen-3 (P. berghei) were unsuccessful, indicating it is essential for asexual blood stage development. CEN-4 forms multi-protein complex with other centrins including CEN-3.\n\n**Paper [6] PMID:22912579**: Mentions PfCEN3 localizes to mitotic spindle poles during schizogony (citing paper [1]).\n\n**Paper [7] PMID:20227667**: \"pfCEN3 plasmid\" here refers to a centromere sequence from chromosome 3 of P. falciparum used as a centromere element in artificial chromosome work — NOT the centrin-3 protein. This is a naming collision within the paper (centromere vs centrin). Skip for protein discoveries.\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2008,\n      \"finding\": \"CEN3 (PF3D7_1027700) protein is expressed in sporozoites, asexual blood forms, gametocytes, and oocysts, and localizes in close proximity to the nucleus of sporozoites and asexual schizonts, with immunofluorescence double-staining with alpha-tubulin indicating association with the parasite centrosome (centriolar plaque).\",\n      \"method\": \"Immunoelectron microscopy and confocal/widefield immunofluorescence microscopy with anti-centrin and anti-alpha-tubulin antibodies in P. falciparum\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct localization by immunofluorescence and immunoelectron microscopy in a single study, but antibody used recognizes multiple centrins and does not exclusively report on CEN3\",\n      \"pmids\": [\"18693242\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CEN3 (PF3D7_1027700) localizes to the extranuclear compartment of the centriolar plaque in P. falciparum blood-stage schizonts, as established using a polyclonal rabbit antibody raised against recombinant PfCen3-6His (PF3D7_1027700.1); this extranuclear position is distinct from the intranuclear compartment where microtubules are nucleated.\",\n      \"method\": \"Super-resolution STED microscopy and correlative light/electron tomography with affinity-purified anti-PfCentrin3 antibody in P. falciparum NF54 strain\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — antibody validated by western blot against recombinant PfCen3-6His, localization confirmed by super-resolution STED and correlative electron tomography, single lab but multiple orthogonal imaging methods\",\n      \"pmids\": [\"34535568\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"NF54\"}]\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CEN3 (PF3D7_1027700) undergoes calcium-inducible liquid-liquid phase separation (LLPS) in vitro: recombinant PfCen3 forms droplets upon Ca2+ addition that display fusion and surface wetting; unlike PfCen1, PfCen3 condensates show partial irreversibility (maturation towards a gel-like state) upon EDTA addition, a process slowed at lower temperature. Mg2+ causes only a reduced response compared to Ca2+, indicating calcium-specificity. The disordered N-terminus and calcium-binding EF-hand motifs are defined as essential features for this condensation.\",\n      \"method\": \"In vitro recombinant protein phase separation assay (6His-tagged and untagged PfCen3 expressed in E. coli), turbidity/light-scattering measurements, live droplet imaging including fusion-event analysis at varying temperatures, EDTA dissolution assays\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with multiple orthogonal quantitative readouts (turbidity, imaging, temperature variation, tagged and untagged protein), single lab but rigorous biochemical controls including calcium specificity and concentration-dependence\",\n      \"pmids\": [\"38150475\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CEN3 (PF3D7_1027700) was identified as a specific co-immunoprecipitation hit with PfCen1-GFP in P. falciparum, indicating a physical interaction between CEN3 and CEN1 at the centriolar plaque; CEN3 co-localizes with the Sfi1-like centriolar plaque protein PfSlp at the extranuclear centriolar plaque as shown by dual-color STED microscopy.\",\n      \"method\": \"Co-immunoprecipitation (PfCen1-GFP pulldown with anti-PfCen3 as readout), reciprocal IP, dual-color STED super-resolution microscopy with anti-PfCen3 antibody in P. falciparum\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — reciprocal Co-IP and super-resolution imaging in single lab, but no cross-linking and quantitative enrichment analysis not performed; interaction corroborated by previously published yeast two-hybrid data cited in abstract\",\n      \"pmids\": [\"37130129\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In P. berghei, deletion of cen-3 (ortholog of PF3D7_1027700) could not be achieved, indicating that CEN3 is likely essential for asexual blood-stage development. Additionally, CEN-3 forms part of a multi-protein complex with other centrins (CEN-1, CEN-2, CEN-4) as shown by complex formation experiments with PbCEN-4.\",\n      \"method\": \"Gene deletion attempts in P. berghei (failed knockout indicating essentiality); multi-protein complex formation analysis with other centrins\",\n      \"journal\": \"Biology open\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — essentiality inferred from failed deletion attempts (single lab), complex formation reported without full mechanistic detail in abstract; consistent with inability to delete cen-1 and cen-2 as well\",\n      \"pmids\": [\"30541825\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Plasmodium berghei\", \"strain\": \"\"}]\n    }\n  ],\n  \"current_model\": \"CEN3 (PF3D7_1027700) is an essential calcium-binding centrin that localizes to the extranuclear compartment of the P. falciparum centriolar plaque, physically interacts with CEN1 and other centrins in a multi-protein complex, and undergoes calcium-induced liquid-liquid phase separation in vitro with partial irreversible condensate maturation, suggesting that Ca2+-regulated biomolecular condensation drives its accumulation at the centrosome during cell division.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CEN3 (PF3D7_1027700) is a calcium-binding centrin that associates with the centrosome-equivalent centriolar plaque of Plasmodium during cell division [#0, #1]. It is expressed across sporozoite, asexual blood, gametocyte, and oocyst stages and localizes in close proximity to the nucleus, specifically to the extranuclear compartment of the centriolar plaque — a position distinct from the intranuclear region where microtubules are nucleated [#0, #1]. At this site CEN3 physically interacts with CEN1 (by reciprocal co-immunoprecipitation) and co-localizes with the Sfi1-like plaque protein PfSlp, and in the P. berghei ortholog it forms part of a multi-protein centrin complex including CEN-1, CEN-2, and CEN-4 [#3, #4]. In P. berghei, cen-3 could not be deleted, indicating the gene is essential for asexual blood-stage development [#4]. Biochemically, recombinant CEN3 undergoes calcium-inducible liquid-liquid phase separation in vitro, forming droplets that fuse and wet surfaces; unlike CEN1, CEN3 condensates mature partially irreversibly toward a gel-like state, a temperature-dependent process, and this behavior depends on its disordered N-terminus and EF-hand calcium-binding motifs, establishing Ca2+-specific condensation as a defining property [#2]. Beyond its centriolar-plaque localization, essentiality, centrin-complex membership, and calcium-driven condensation, the precise cellular function of CEN3 in centriolar plaque biology has not been mechanistically resolved in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 2008,\n      \"claim\": \"Established that CEN3 is a broadly expressed centrin positioned at the parasite centrosome, placing it at the machinery of cell division.\",\n      \"evidence\": \"Immunoelectron and confocal/widefield immunofluorescence microscopy with anti-centrin and anti-alpha-tubulin antibodies in P. falciparum\",\n      \"pmids\": [\"18693242\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Antibody recognizes multiple centrins and does not exclusively report CEN3\",\n        \"No functional role assigned beyond spatial association with the centrosome\",\n        \"Sub-compartment within the centriolar plaque not resolved\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Tested whether CEN3 is dispensable and whether it acts alone; the ortholog proved essential and was shown to sit within a multi-centrin complex.\",\n      \"evidence\": \"Failed cen-3 gene deletion (essentiality inference) and multi-protein complex formation analysis with PbCEN-4 in P. berghei\",\n      \"pmids\": [\"30541825\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Essentiality inferred from failed knockout rather than conditional depletion\",\n        \"Stoichiometry and architecture of the centrin complex not defined\",\n        \"Direct versus indirect nature of CEN3 contacts within the complex unresolved\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium berghei\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Resolved where within the centriolar plaque CEN3 acts, distinguishing its extranuclear position from the intranuclear microtubule-nucleating compartment.\",\n      \"evidence\": \"Super-resolution STED microscopy and correlative light/electron tomography with affinity-purified anti-PfCentrin3 antibody in P. falciparum NF54\",\n      \"pmids\": [\"34535568\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Functional consequence of extranuclear localization not established\",\n        \"Dynamics of CEN3 recruitment during the cell cycle not tracked\",\n        \"Molecular tethers holding CEN3 in the extranuclear compartment unknown\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"NF54\"}]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined CEN3's physical partners at the plaque, connecting it to CEN1 and the Sfi1-like protein PfSlp.\",\n      \"evidence\": \"Reciprocal co-immunoprecipitation (PfCen1-GFP pulldown) and dual-color STED super-resolution microscopy in P. falciparum\",\n      \"pmids\": [\"37130129\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No cross-linking or quantitative enrichment analysis performed\",\n        \"Whether the CEN3-CEN1 interaction is direct not shown\",\n        \"Functional role of the CEN3-PfSlp co-localization not tested\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Provided a biophysical mechanism for CEN3 accumulation by showing calcium-specific, EF-hand- and N-terminus-dependent phase separation, and distinguished CEN3 from CEN1 by its partial condensate irreversibility.\",\n      \"evidence\": \"In vitro reconstitution with recombinant tagged and untagged PfCen3, turbidity/light-scattering, live droplet fusion imaging at varying temperatures, and EDTA dissolution assays\",\n      \"pmids\": [\"38150475\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Phase separation demonstrated in vitro only, not confirmed at the centriolar plaque in vivo\",\n        \"Physiological consequence of irreversible condensate maturation unknown\",\n        \"Whether CEN3 condensation recruits or scaffolds its complex partners not tested\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How Ca2+-driven CEN3 condensation is coupled to its essential role in centriolar plaque function and parasite division remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No conditional depletion linking loss of CEN3 to a specific division defect\",\n        \"In vitro phase separation not yet connected to in vivo condensate formation\",\n        \"Direct molecular activity of CEN3 within the plaque undefined\"\n      ],\n      \"attribution\": [{\"species\": \"Plasmodium falciparum\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005509\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005813\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0051301\", \"supporting_discovery_ids\": [1, 4]}\n    ],\n    \"complexes\": [\n      \"Centrin multi-protein complex (CEN-1, CEN-2, CEN-3, CEN-4) at the centriolar plaque\"\n    ],\n    \"partners\": [\n      \"CEN1 (PfCen1)\",\n      \"PfSlp (Sfi1-like centriolar plaque protein)\",\n      \"PbCEN-1, PbCEN-2, PbCEN-4 (in P. berghei)\"\n    ],\n    \"other_free_text\": [\n      \"Undergoes calcium-inducible liquid-liquid phase separation in vitro; condensates mature partially irreversibly toward a gel-like state, distinguishing CEN3 from CEN1\",\n      \"Condensation depends on the disordered N-terminus and EF-hand calcium-binding motifs; calcium-specific (Mg2+ gives reduced response)\",\n      \"Localizes to the extranuclear compartment of the centriolar plaque, distinct from the intranuclear microtubule-nucleating region\",\n      \"Essential in P. berghei asexual blood stage (cen-3 deletion could not be achieved)\",\n      \"Expressed in sporozoites, asexual blood forms, gametocytes, and oocysts\"\n    ]\n  }\n}","audit_flag":null,"strains":[{"strain":"Pf3D7","gene_id":"PF3D7_1027700","name":null,"product":"centrin-3","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PbANKA","gene_id":"PBANKA_0511800","name":null,"product":"centrin-3, putative","uniprot_acc":null,"chromosome":null,"start_pos":null},{"strain":"PkH","gene_id":"PKNH_0612100","name":null,"product":"centrin-3, putative","uniprot_acc":null,"chromosome":null,"start_pos":null}],"gene_symbol":"CEN3","evaluation":null}