{"id":"601fc607-870d-5caa-8f10-1cad2ab9b98f","stable_key":"f992b796-377d-53bf-a39b-7e5d41dbe194:b5-met-pdk2-coalation-inhibition","predicate":"inhibits_after_coalation","statement":"In-vitro CoAlation of immunoprecipitated recombinant PDK2 reduced its phosphorylation of PDH E1, and DTT restored kinase activity.","claim_class":"mechanistic","status":"source_derived_draft","evidence_grade":"ungraded","direction":"negative","is_public":true,"mechanism_event_id":"a28f47d0-d453-5c04-8486-5209364d0eb6","mechanism_event_label":"Adding CoA through a disulfide bond reversibly inhibited the kinase that regulates pyruvate dehydrogenase.","subject":{"id":"0d4dcd7b-d981-56ba-9172-221c17faaa3d","slug":"coa-disulfide","display_name":"Coenzyme A disulfide","entity_type_key":"small_molecule"},"object":{"id":"ec53c923-a38e-57ce-83a4-dbd4a3be7038","slug":"pdk2","display_name":"Human pyruvate dehydrogenase kinase 2 / PDK2","entity_type_key":"protein"},"evidence_count":1,"mechanism_event":{"id":"a28f47d0-d453-5c04-8486-5209364d0eb6","stable_key":"f992b796-377d-53bf-a39b-7e5d41dbe194:b5-met-pdk2-coalation-inhibition-event","event_type":"biochemical_relationship","label":"Adding CoA through a disulfide bond reversibly inhibited the kinase that regulates pyruvate dehydrogenase.","description":"In-vitro CoAlation of immunoprecipitated recombinant PDK2 reduced its phosphorylation of PDH E1, and DTT restored kinase activity.","status":"provisional","compartment":null,"participants":[{"entity":{"id":"bd80a874-773e-557d-9f47-6486ee2efe04","slug":"pdk2-coalated","display_name":"Human CoAlated PDK2","entity_type_key":"protein_state"},"role":"modified_enzyme_state","stoichiometry":null,"state_label":"","sequence_order":0,"notes":""},{"entity":{"id":"a334fc3d-c73c-5733-8f8c-c028361da270","slug":"porcine-heart-pyruvate-dehydrogenase-complex","display_name":"Porcine-heart pyruvate dehydrogenase complex","entity_type_key":"protein_complex"},"role":"kinase_substrate","stoichiometry":null,"state_label":"","sequence_order":1,"notes":"Radioactive phosphate incorporation into its E1 subunit was measured."},{"entity":{"id":"4faa6456-aff8-59eb-9e3f-3326a436e401","slug":"coenzyme-a","display_name":"Coenzyme A","entity_type_key":"small_molecule"},"role":"modifying_moiety","stoichiometry":null,"state_label":"","sequence_order":2,"notes":""},{"entity":{"id":"0d4dcd7b-d981-56ba-9172-221c17faaa3d","slug":"coa-disulfide","display_name":"Coenzyme A disulfide","entity_type_key":"small_molecule"},"role":"subject","stoichiometry":null,"state_label":"","sequence_order":3,"notes":""},{"entity":{"id":"ec53c923-a38e-57ce-83a4-dbd4a3be7038","slug":"pdk2","display_name":"Human pyruvate dehydrogenase kinase 2 / PDK2","entity_type_key":"protein"},"role":"target","stoichiometry":null,"state_label":"","sequence_order":4,"notes":""}]},"contexts":[{"dimension":"cross_nutrient","value_text":"false","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"evidence_location","value_text":"Full text lines 33, 61 and 141; Fig. 6D","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"experimental_model","value_text":"FLAG-PDK2 expressed in HEK293 cells, immunoprecipitated and assayed with porcine-heart PDH","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"exposure","value_text":"1 mM CoA disulfide preincubation for 20 minutes; PDH phosphorylation assay with/without 2 mM DTT.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"limitations","value_text":"CoA disulfide is distinct from reduced CoA. The assay does not establish net mitochondrial PDH flux or a supplementation effect in vivo. This experiment does not establish a dietary pantothenate threshold or benefit from B5 supplementation.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"nutrient_topic","value_text":"Pantothenic acid (vitamin B5) research collection; topical membership is not evidence of a direct dietary effect.","comparator":null,"unit":null,"notes":"","entity":{"slug":"pantothenate","display_name":"Pantothenate (vitamin B5)","entity_type_key":"small_molecule"}},{"dimension":"organism","value_text":"Human PDK2; Sus scrofa PDH assay substrate","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"plain_language","value_text":"Adding CoA through a disulfide bond reversibly inhibited the kinase that regulates pyruvate dehydrogenase.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"primary_references","value_text":"[b5-met-coalation2017] Protein CoAlation: a redox-regulated protein modification by coenzyme A in mammalian cells. (2017). https://pubmed.ncbi.nlm.nih.gov/28341808/ DOI: 10.1042/bcj20170129","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"tissue_or_cell_type","value_text":"Cell-free kinase assay","comparator":null,"unit":null,"notes":"","entity":null}],"evidence":[{"id":"80c85006-8e94-5f5c-b9d4-e5466efebff6","evidence_kind":"source_excerpt","locator":"Lines 1089-1101","start_line":1089,"end_line":1101,"excerpt":"### b5-met-pdk2-coalation-inhibition\nIn-vitro CoAlation of immunoprecipitated recombinant PDK2 reduced its phosphorylation of PDH E1, and DTT restored kinase activity.\nCondition category: normal\nnutrient_topic: Pantothenic acid (vitamin B5) research collection; topical membership is not evidence of a direct dietary effect.\nplain_language: Adding CoA through a disulfide bond reversibly inhibited the kinase that regulates pyruvate dehydrogenase.\norganism: Human PDK2; Sus scrofa PDH assay substrate\ntissue_or_cell_type: Cell-free kinase assay\nexperimental_model: FLAG-PDK2 expressed in HEK293 cells, immunoprecipitated and assayed with porcine-heart PDH\nlimitations: CoA disulfide is distinct from reduced CoA. The assay does not establish net mitochondrial PDH flux or a supplementation effect in vivo. This experiment does not establish a dietary pantothenate threshold or benefit from B5 supplementation.\nexposure: 1 mM CoA disulfide preincubation for 20 minutes; PDH phosphorylation assay with/without 2 mM DTT.\ncross_nutrient: false\nevidence_location: Full text lines 33, 61 and 141; Fig. 6D\n[b5-met-coalation2017] Protein CoAlation: a redox-regulated protein modification by coenzyme A in mammalian cells. (2017). https://pubmed.ncbi.nlm.nih.gov/28341808/ DOI: 10.1042/bcj20170129","model_system":"FLAG-PDK2 expressed in HEK293 cells, immunoprecipitated and assayed with porcine-heart PDH","directness":"author_interpretation","verification_status":"source_derived_draft","notes":"Exact curation-document quotation, not publisher quotation. Study references: [b5-met-coalation2017] Protein CoAlation: a redox-regulated protein modification by coenzyme A in mammalian cells. (2017). https://pubmed.ncbi.nlm.nih.gov/28341808/ DOI: 10.1042/bcj20170129","relationship":"supports","weight":1.0,"link_notes":"","source":{"id":"0716b500-dd0b-5425-b9d0-d88bc9c09e86","stable_key":"import-f992b796-377d-53bf-a39b-7e5d41dbe194","title":"Pantothenic acid (vitamin B5): coenzyme A, deficiency and nutrient interactions (2026-09-17)","document_type":"imported_text","citation_label":"AI-assisted literature curation; primary study URLs and scope retained in the document and extraction. Not publisher full text.","file_path":"","sha256":"b5a49f5a2373ca6064c9a4e014e052dad2f6f199a82bb09b26b82b831c36110b","revision_id":"78f3e793-f084-5ee2-8703-b661c4b650bf","review_status":"unverified_draft","notes":""}}],"relations":[],"conflicts":[],"corrections":[],"research":null}