{"id":"447af834-ae9a-54ea-ab2a-68fb654a7c48","stable_key":"e0ea2d6a-7429-5e9f-b774-41e5e3288da3:e-sig-nbd-pkc-binding","predicate":"binds","statement":"NBD-alpha-tocopherol bound recombinant human PKC alpha in solution: fluorescence increased at 1 µM probe, and 5 µM was used in competition assays with 0.1 µM enzyme.","claim_class":"mechanistic","status":"source_derived_draft","evidence_grade":"ungraded","direction":"positive","is_public":true,"mechanism_event_id":"38fc7639-f298-5bbf-b92d-47e5f1e1d28c","mechanism_event_label":"A fluorescently tagged vitamin E probe associated directly with purified PKC alpha.","subject":{"id":"0671a181-7c20-55b1-832b-8ea7da35052e","slug":"nbd-alpha-tocopherol","display_name":"NBD-alpha-tocopherol","entity_type_key":"small_molecule"},"object":{"id":"d4c19e4b-d83b-5cda-8c7d-041d7fe6ada0","slug":"prkca","display_name":"Protein kinase C alpha (PRKCA)","entity_type_key":"protein"},"evidence_count":1,"mechanism_event":{"id":"38fc7639-f298-5bbf-b92d-47e5f1e1d28c","stable_key":"e0ea2d6a-7429-5e9f-b774-41e5e3288da3:e-sig-nbd-pkc-binding-event","event_type":"biochemical_relationship","label":"A fluorescently tagged vitamin E probe associated directly with purified PKC alpha.","description":"NBD-alpha-tocopherol bound recombinant human PKC alpha in solution: fluorescence increased at 1 µM probe, and 5 µM was used in competition assays with 0.1 µM enzyme.","status":"provisional","compartment":null,"participants":[{"entity":{"id":"0671a181-7c20-55b1-832b-8ea7da35052e","slug":"nbd-alpha-tocopherol","display_name":"NBD-alpha-tocopherol","entity_type_key":"small_molecule"},"role":"subject","stoichiometry":null,"state_label":"","sequence_order":0,"notes":""},{"entity":{"id":"d4c19e4b-d83b-5cda-8c7d-041d7fe6ada0","slug":"prkca","display_name":"Protein kinase C alpha (PRKCA)","entity_type_key":"protein"},"role":"target","stoichiometry":null,"state_label":"","sequence_order":1,"notes":""}]},"contexts":[{"dimension":"cross_nutrient","value_text":"false","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"experimental_model","value_text":"Fluorescence ligand binding in solution","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"exposure","value_text":"0.1 µM PKC alpha; 1–5 µM NBD-alpha-tocopherol; 30 min at room temperature; 1% ethanol buffer.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"limitations","value_text":"The fluorophore reports a hydrophobic environment. Competition controls support specific binding, but the labeled probe is distinct from native tocopherol.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"nutrient_topic","value_text":"Vitamin E research collection; topical membership is not evidence of a direct dietary effect.","comparator":null,"unit":null,"notes":"","entity":{"slug":"vitamin-e","display_name":"Vitamin E","entity_type_key":"chemical_species"}},{"dimension":"organism","value_text":"Homo sapiens","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"plain_language","value_text":"A fluorescently tagged vitamin E probe associated directly with purified PKC alpha.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"primary_references","value_text":"[mccary2012] Vitamin E isoforms directly bind PKCα and differentially regulate activation of PKCα. (2012). https://pubmed.ncbi.nlm.nih.gov/21933153/ DOI: 10.1042/bj20111318","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"tissue_or_cell_type","value_text":"Cell-free enzyme","comparator":null,"unit":null,"notes":"","entity":null}],"evidence":[{"id":"b5ac034c-4fa0-51fd-b2f4-0613aa217463","evidence_kind":"source_excerpt","locator":"Lines 972-983","start_line":972,"end_line":983,"excerpt":"### e-sig-nbd-pkc-binding\nNBD-alpha-tocopherol bound recombinant human PKC alpha in solution: fluorescence increased at 1 µM probe, and 5 µM was used in competition assays with 0.1 µM enzyme.\nCondition category: normal\nnutrient_topic: Vitamin E research collection; topical membership is not evidence of a direct dietary effect.\nplain_language: A fluorescently tagged vitamin E probe associated directly with purified PKC alpha.\norganism: Homo sapiens\ntissue_or_cell_type: Cell-free enzyme\nexperimental_model: Fluorescence ligand binding in solution\nlimitations: The fluorophore reports a hydrophobic environment. Competition controls support specific binding, but the labeled probe is distinct from native tocopherol.\nexposure: 0.1 µM PKC alpha; 1–5 µM NBD-alpha-tocopherol; 30 min at room temperature; 1% ethanol buffer.\ncross_nutrient: false\n[mccary2012] Vitamin E isoforms directly bind PKCα and differentially regulate activation of PKCα. (2012). https://pubmed.ncbi.nlm.nih.gov/21933153/ DOI: 10.1042/bj20111318","model_system":"Fluorescence ligand binding in solution","directness":"author_interpretation","verification_status":"source_derived_draft","notes":"Exact curation-document quotation, not publisher quotation. Study references: [mccary2012] Vitamin E isoforms directly bind PKCα and differentially regulate activation of PKCα. (2012). https://pubmed.ncbi.nlm.nih.gov/21933153/ DOI: 10.1042/bj20111318","relationship":"supports","weight":1.0,"link_notes":"","source":{"id":"3ac41dba-4727-5116-a556-ac3bbb0dc41c","stable_key":"import-e0ea2d6a-7429-5e9f-b774-41e5e3288da3","title":"Vitamin E: transport, membrane protection 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":"a90b6565e092c6a2da07f74cf758792c2cf63b40746260c8ff12bd64322c36f4","revision_id":"956793d1-23e4-5397-9291-25c1732a8e02","review_status":"unverified_draft","notes":""}}],"relations":[],"conflicts":[],"corrections":[],"research":null}