{"id":"3f06da15-a304-5183-9010-64a9417a395a","stable_key":"3c852f36-7951-5fd9-9013-d771a8d16d8a:vae-rgr-rdh10-coupled-retinol","predicate":"cooperates_with","statement":"HEK293T cells coexpressing bovine RGR and RDH10 generated more 11-cis-retinol from all-trans-retinol during illumination than cells expressing either protein alone.","claim_class":"mechanistic","status":"source_derived_draft","evidence_grade":"ungraded","direction":"positive","is_public":true,"mechanism_event_id":"c781ef63-48c0-5076-96d1-1a53226d2412","mechanism_event_label":"The two proteins supported light-dependent retinol recycling together in cultured cells.","subject":{"id":"205eda81-4688-5fa3-a08c-e94d0c1cbf5c","slug":"rgr","display_name":"Retinal G protein-coupled receptor / RGR opsin","entity_type_key":"protein"},"object":{"id":"89b4b04a-7637-56f4-ac55-501ad31d1f61","slug":"rdh10","display_name":"Retinol dehydrogenase 10 / RDH10","entity_type_key":"protein"},"evidence_count":1,"mechanism_event":{"id":"c781ef63-48c0-5076-96d1-1a53226d2412","stable_key":"3c852f36-7951-5fd9-9013-d771a8d16d8a:vae-rgr-rdh10-coupled-retinol-event","event_type":"biochemical_relationship","label":"The two proteins supported light-dependent retinol recycling together in cultured cells.","description":"HEK293T cells coexpressing bovine RGR and RDH10 generated more 11-cis-retinol from all-trans-retinol during illumination than cells expressing either protein alone.","status":"provisional","compartment":null,"participants":[{"entity":{"id":"205eda81-4688-5fa3-a08c-e94d0c1cbf5c","slug":"rgr","display_name":"Retinal G protein-coupled receptor / RGR opsin","entity_type_key":"protein"},"role":"photoisomerase","stoichiometry":null,"state_label":"","sequence_order":0,"notes":""},{"entity":{"id":"89b4b04a-7637-56f4-ac55-501ad31d1f61","slug":"rdh10","display_name":"Retinol dehydrogenase 10 / RDH10","entity_type_key":"protein"},"role":"coexpressed oxidoreductase","stoichiometry":null,"state_label":"","sequence_order":1,"notes":""},{"entity":{"id":"148c57d8-c658-5769-89e8-1194cef2a782","slug":"retinol","display_name":"All-trans-retinol","entity_type_key":"small_molecule"},"role":"substrate","stoichiometry":null,"state_label":"","sequence_order":2,"notes":""},{"entity":{"id":"4a978267-139b-518d-bdc6-e16cda6f5225","slug":"11-cis-retinol","display_name":"11-cis-retinol","entity_type_key":"small_molecule"},"role":"product","stoichiometry":null,"state_label":"","sequence_order":3,"notes":""},{"entity":{"id":"4aba2a5e-8d06-5304-bb01-c0402b225a94","slug":"nadph","display_name":"NADPH","entity_type_key":"small_molecule"},"role":"added assay reagent","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":"Figure 1A; STAR Methods: Expression of RGR and RDH10","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"experimental_model","value_text":"Recombinant bovine RGR/RDH10 coexpression","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"exposure","value_text":"30-minute illumination; 5 micromolar all-trans-retinol; assay medium contained 250 micromolar NADPH.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"limitations","value_text":"Coupled output does not prove every proposed intermediate reaction or RDH10 necessity in intact retina. Added NADPH is an assay component, not a demonstrated dietary requirement.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"nutrient","value_text":"Vitamin A","comparator":null,"unit":null,"notes":"","entity":{"slug":"vitamin-a","display_name":"Vitamin A","entity_type_key":"chemical_species"}},{"dimension":"nutrient_topic","value_text":"Vitamin A expansion research collection; topical membership is not evidence of a direct dietary effect.","comparator":null,"unit":null,"notes":"","entity":{"slug":"vitamin-a","display_name":"Vitamin A","entity_type_key":"chemical_species"}},{"dimension":"organism","value_text":"Bos taurus proteins in Homo sapiens cells","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"plain_language","value_text":"The two proteins supported light-dependent retinol recycling together in cultured cells.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"primary_references","value_text":"[vav-morshedian2019] Light-Driven Regeneration of Cone Visual Pigments through a Mechanism Involving RGR Opsin in Müller Glial Cells. (2019). https://pubmed.ncbi.nlm.nih.gov/31056353/ DOI: 10.1016/j.neuron.2019.04.004","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"tissue_or_cell_type","value_text":"HEK293T culture","comparator":null,"unit":null,"notes":"","entity":null}],"evidence":[{"id":"002c9a37-072d-5379-8f50-4a74e6da04e3","evidence_kind":"source_excerpt","locator":"Lines 468-481","start_line":468,"end_line":481,"excerpt":"### vae-rgr-rdh10-coupled-retinol\nHEK293T cells coexpressing bovine RGR and RDH10 generated more 11-cis-retinol from all-trans-retinol during illumination than cells expressing either protein alone.\nCondition category: normal\nnutrient_topic: Vitamin A expansion research collection; topical membership is not evidence of a direct dietary effect.\nplain_language: The two proteins supported light-dependent retinol recycling together in cultured cells.\norganism: Bos taurus proteins in Homo sapiens cells\ntissue_or_cell_type: HEK293T culture\nexperimental_model: Recombinant bovine RGR/RDH10 coexpression\nlimitations: Coupled output does not prove every proposed intermediate reaction or RDH10 necessity in intact retina. Added NADPH is an assay component, not a demonstrated dietary requirement.\nexposure: 30-minute illumination; 5 micromolar all-trans-retinol; assay medium contained 250 micromolar NADPH.\ncross_nutrient: false\nevidence_location: Figure 1A; STAR Methods: Expression of RGR and RDH10\nnutrient: Vitamin A\n[vav-morshedian2019] Light-Driven Regeneration of Cone Visual Pigments through a Mechanism Involving RGR Opsin in Müller Glial Cells. (2019). https://pubmed.ncbi.nlm.nih.gov/31056353/ DOI: 10.1016/j.neuron.2019.04.004","model_system":"Recombinant bovine RGR/RDH10 coexpression","directness":"author_interpretation","verification_status":"source_derived_draft","notes":"Exact curation-document quotation, not publisher quotation. Study references: [vav-morshedian2019] Light-Driven Regeneration of Cone Visual Pigments through a Mechanism Involving RGR Opsin in Müller Glial Cells. 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