{"id":"6a75665e-e24d-589c-a57c-0718dba7071a","stable_key":"220adaab-10d8-5c4e-a3f5-a48700ea794d:agmatine-sulfate-rat-alp-metal","predicate":"is_required_for","statement":"Rat ALP catalytic activity required manganese; D217A and specified multi-residue substitutions abolished measured agmatinase activity.","claim_class":"mechanistic","status":"source_derived_draft","evidence_grade":"ungraded","direction":"context_dependent","is_public":true,"mechanism_event_id":"ee629fd7-646d-525a-9728-8ddb4bc46011","mechanism_event_label":"The metal-dependent catalytic machinery is a separate requirement from substrate supply.","subject":{"id":"a8082b11-c484-5792-bb81-48e8e699cbe4","slug":"manganese-ion","display_name":"Mn2+","entity_type_key":"ion"},"object":{"id":"a44bcaf9-a7c7-5d42-a850-6eb7b4fae03c","slug":"rat-agmatinase-like-protein","display_name":"Rat brain agmatinase-like protein / ALP experimental preparation","entity_type_key":"protein"},"evidence_count":1,"mechanism_event":{"id":"ee629fd7-646d-525a-9728-8ddb4bc46011","stable_key":"220adaab-10d8-5c4e-a3f5-a48700ea794d:agmatine-sulfate-rat-alp-metal-event","event_type":"observed_relationship","label":"The metal-dependent catalytic machinery is a separate requirement from substrate supply.","description":"Rat ALP catalytic activity required manganese; D217A and specified multi-residue substitutions abolished measured agmatinase activity.","status":"provisional","compartment":null,"participants":[{"entity":{"id":"a8082b11-c484-5792-bb81-48e8e699cbe4","slug":"manganese-ion","display_name":"Mn2+","entity_type_key":"ion"},"role":"subject","stoichiometry":null,"state_label":"","sequence_order":0,"notes":""},{"entity":{"id":"a44bcaf9-a7c7-5d42-a850-6eb7b4fae03c","slug":"rat-agmatinase-like-protein","display_name":"Rat brain agmatinase-like protein / ALP experimental preparation","entity_type_key":"protein"},"role":"target","stoichiometry":null,"state_label":"","sequence_order":1,"notes":""},{"entity":{"id":"ae2dddf4-ce04-57e3-8a06-7ed9fdc24554","slug":"agmatine-sulfate","display_name":"Agmatine Sulfate","entity_type_key":"small_molecule"},"role":"context_participant","stoichiometry":null,"state_label":"","sequence_order":2,"notes":""},{"entity":{"id":"e2f83eb4-99ed-5050-87d3-3e4701d05c1a","slug":"agmatine","display_name":"Agmatine","entity_type_key":"small_molecule"},"role":"context_participant","stoichiometry":null,"state_label":"","sequence_order":3,"notes":""},{"entity":{"id":"c71bf622-f14b-5314-b1a5-dfa002d2d8ff","slug":"putrescine","display_name":"Putrescine","entity_type_key":"small_molecule"},"role":"context_participant","stoichiometry":null,"state_label":"","sequence_order":4,"notes":""}]},"contexts":[{"dimension":"availability_state","value_text":"machinery_impairment","comparator":null,"unit":null,"notes":"Imported condition classification; unverified.","entity":null},{"dimension":"evidence_access","value_text":"Primary abstract","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"experimental_model","value_text":"Recombinant rat protein, manganese activation and mutagenesis.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"limitations","value_text":"Mutations support residue importance but do not by themselves prove every proposed metal-coordination geometry.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"nutrient_topic","value_text":"Agmatine Sulfate collection; species, compartment, exposure, co-substrates and manipulation remain explicit.","comparator":null,"unit":null,"notes":"","entity":{"slug":"agmatine-sulfate","display_name":"Agmatine Sulfate","entity_type_key":"small_molecule"}},{"dimension":"plain_language","value_text":"The metal-dependent catalytic machinery is a separate requirement from substrate supply.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"primary_references","value_text":"Insights into the Mn2+ Binding Site in the Agmatinase-Like Protein (ALP): A Critical Enzyme for the Regulation of Agmatine Levels in Mammals. · 2020 · https://pubmed.ncbi.nlm.nih.gov/32531922/ · DOI 10.3390/ijms21114132","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"trigger_kind","value_text":"machinery_impairment","comparator":null,"unit":null,"notes":"Imported condition classification; unverified.","entity":null}],"evidence":[{"id":"13a912da-e365-5f54-888d-0ceeb92329cc","evidence_kind":"source_excerpt","locator":"Lines 156-162","start_line":156,"end_line":162,"excerpt":"## agmatine-sulfate-rat-alp-metal\nThe metal-dependent catalytic machinery is a separate requirement from substrate supply.\nRat ALP catalytic activity required manganese; D217A and specified multi-residue substitutions abolished measured agmatinase activity.\nModel: Recombinant rat protein, manganese activation and mutagenesis.\nLimitations: Mutations support residue importance but do not by themselves prove every proposed metal-coordination geometry.\nEvidence access: Primary abstract\nInsights into the Mn2+ Binding Site in the Agmatinase-Like Protein (ALP): A Critical Enzyme for the Regulation of Agmatine Levels in Mammals. · 2020 · https://pubmed.ncbi.nlm.nih.gov/32531922/ · DOI 10.3390/ijms21114132","model_system":"Recombinant rat protein, manganese activation and mutagenesis.","directness":"reported_statement","verification_status":"source_derived_draft","notes":"Original curation paraphrase; 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