{"id":"a2d4e944-edc8-5a03-9021-7ef22ab52e9d","stable_key":"6d38d43e-01e4-5641-93be-65654271e242:zinc-enz-sod1-zinc-binding","predicate":"promotes_formation_of","statement":"Adding 10 µM ZnSO4 during human SOD1 expression in HEK293T cells promoted site-selective binding of one zinc ion per SOD1 subunit and the zinc-containing dimer.","claim_class":"mechanistic","status":"source_derived_draft","evidence_grade":"ungraded","direction":"positive","is_public":true,"mechanism_event_id":"9eb6de75-7e5e-58f5-b6da-d42d29cdcbc9","mechanism_event_label":"Zinc binding helps the cellular SOD1 protein reach a dimeric maturation intermediate.","subject":{"id":"49c806c2-7041-5020-8b3b-fa04ffa122ac","slug":"zinc-ion","display_name":"Zinc(II) ion","entity_type_key":"ion"},"object":{"id":"a2f18cf0-b9dd-5ba2-962f-e76ac8dbaa2c","slug":"sod1-zinc-bound-reduced","display_name":"Zinc-bound disulfide-reduced human SOD1","entity_type_key":"protein_state"},"evidence_count":1,"mechanism_event":{"id":"9eb6de75-7e5e-58f5-b6da-d42d29cdcbc9","stable_key":"6d38d43e-01e4-5641-93be-65654271e242:zinc-enz-sod1-zinc-binding-event","event_type":"biochemical_relationship","label":"Zinc binding helps the cellular SOD1 protein reach a dimeric maturation intermediate.","description":"Adding 10 µM ZnSO4 during human SOD1 expression in HEK293T cells promoted site-selective binding of one zinc ion per SOD1 subunit and the zinc-containing dimer.","status":"provisional","compartment":null,"participants":[{"entity":{"id":"19f2bdc0-4888-57d5-860f-119d5d478eb4","slug":"sod1","display_name":"Human copper-zinc superoxide dismutase / SOD1","entity_type_key":"protein"},"role":"precursor","stoichiometry":null,"state_label":"","sequence_order":0,"notes":""},{"entity":{"id":"9f823444-e2dd-5e16-9440-1bf2f62e8bdf","slug":"zinc-sulfate","display_name":"Zinc sulfate","entity_type_key":"chemical_species"},"role":"medium_addition","stoichiometry":null,"state_label":"","sequence_order":1,"notes":""},{"entity":{"id":"49c806c2-7041-5020-8b3b-fa04ffa122ac","slug":"zinc-ion","display_name":"Zinc(II) ion","entity_type_key":"ion"},"role":"subject","stoichiometry":null,"state_label":"","sequence_order":2,"notes":""},{"entity":{"id":"a2f18cf0-b9dd-5ba2-962f-e76ac8dbaa2c","slug":"sod1-zinc-bound-reduced","display_name":"Zinc-bound disulfide-reduced human SOD1","entity_type_key":"protein_state"},"role":"target","stoichiometry":null,"state_label":"","sequence_order":3,"notes":""}]},"contexts":[{"dimension":"cross_nutrient","value_text":"false","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"experimental_model","value_text":"Transient expression of human SOD1 and CCS in human HEK293T cells; in-cell NMR","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"exposure","value_text":"10 µM ZnSO4 immediately after transfection; 48 h expression before later copper manipulations.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"limitations","value_text":"HEK293T cells overexpressed SOD1/CCS above endogenous levels; these medium concentrations are experimental exposures, not dietary targets. Copper transfer and disulfide oxidation need not occur in a fixed coupled step in every cellular context.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"nutrient_topic","value_text":"Zinc research collection; topical membership is not evidence of a direct dietary effect.","comparator":null,"unit":null,"notes":"","entity":{"slug":"zinc","display_name":"Zinc","entity_type_key":"nutrient_element"}},{"dimension":"organism","value_text":"Homo sapiens","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"plain_language","value_text":"Zinc binding helps the cellular SOD1 protein reach a dimeric maturation intermediate.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"primary_references","value_text":"[zinc-enz-sod1-live2013] Atomic-resolution monitoring of protein maturation in live human cells by NMR. (2013). https://pubmed.ncbi.nlm.nih.gov/23455544/ DOI: 10.1038/nchembio.1202","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"tissue_or_cell_type","value_text":"Human HEK293T cytoplasm","comparator":null,"unit":null,"notes":"","entity":null}],"evidence":[{"id":"52504a30-008e-5134-8684-8d13c4edd637","evidence_kind":"source_excerpt","locator":"Lines 729-740","start_line":729,"end_line":740,"excerpt":"### zinc-enz-sod1-zinc-binding\nAdding 10 µM ZnSO4 during human SOD1 expression in HEK293T cells promoted site-selective binding of one zinc ion per SOD1 subunit and the zinc-containing dimer.\nCondition category: normal\nnutrient_topic: Zinc research collection; topical membership is not evidence of a direct dietary effect.\nplain_language: Zinc binding helps the cellular SOD1 protein reach a dimeric maturation intermediate.\norganism: Homo sapiens\ntissue_or_cell_type: Human HEK293T cytoplasm\nexperimental_model: Transient expression of human SOD1 and CCS in human HEK293T cells; in-cell NMR\nlimitations: HEK293T cells overexpressed SOD1/CCS above endogenous levels; these medium concentrations are experimental exposures, not dietary targets. Copper transfer and disulfide oxidation need not occur in a fixed coupled step in every cellular context.\nexposure: 10 µM ZnSO4 immediately after transfection; 48 h expression before later copper manipulations.\ncross_nutrient: false\n[zinc-enz-sod1-live2013] Atomic-resolution monitoring of protein maturation in live human cells by NMR. (2013). https://pubmed.ncbi.nlm.nih.gov/23455544/ DOI: 10.1038/nchembio.1202","model_system":"Transient expression of human SOD1 and CCS in human HEK293T cells; in-cell NMR","directness":"author_interpretation","verification_status":"source_derived_draft","notes":"Exact curation-document quotation, not publisher quotation. Study references: [zinc-enz-sod1-live2013] Atomic-resolution monitoring of protein maturation in live human cells by NMR. 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