{"id":"e6dad842-8439-5e3b-bc89-8b1f3d048a0e","stable_key":"7fc92b9e-9cbf-556e-8719-6b5244625250:b12-abs-amn-trafficking-rat-construct","predicate":"enables-surface-targeting-of","statement":"Human AMN coexpression allowed rat mini-cubilin to reach the plasma membrane in HEK293T and other tested renal/intestinal cell models, whereas mini-cubilin alone remained intracellular.","claim_class":"mechanistic","status":"source_derived_draft","evidence_grade":"ungraded","direction":"positive","is_public":true,"mechanism_event_id":"032d9eff-83ac-5278-aa12-6e5242fcbd90","mechanism_event_label":"Amnionless helped the experimental receptor reach the cell surface.","subject":{"id":"bbd594ac-365b-507f-954d-20a0df38312d","slug":"amn","display_name":"Human amnionless / AMN","entity_type_key":"protein"},"object":{"id":"cca4766f-3ab0-57ea-8cb1-117af937c808","slug":"rat-mini-cubilin","display_name":"Rat N-terminal mini-cubilin construct","entity_type_key":"protein_state"},"evidence_count":1,"mechanism_event":{"id":"032d9eff-83ac-5278-aa12-6e5242fcbd90","stable_key":"7fc92b9e-9cbf-556e-8719-6b5244625250:b12-abs-amn-trafficking-rat-construct-event","event_type":"biochemical_relationship","label":"Amnionless helped the experimental receptor reach the cell surface.","description":"Human AMN coexpression allowed rat mini-cubilin to reach the plasma membrane in HEK293T and other tested renal/intestinal cell models, whereas mini-cubilin alone remained intracellular.","status":"provisional","compartment":null,"participants":[{"entity":{"id":"b6d9b937-895d-5e0a-8cc5-0454a0b7d373","slug":"vitamin-b12","display_name":"Vitamin B12 (cobalamins)","entity_type_key":"chemical_species"},"role":"nutrient","stoichiometry":null,"state_label":"","sequence_order":0,"notes":""},{"entity":{"id":"6d2228b6-1b1b-565e-975b-0f3bf0d46b47","slug":"rat-cubn","display_name":"Rat cubilin / Cubn","entity_type_key":"protein"},"role":"parent-protein","stoichiometry":null,"state_label":"","sequence_order":1,"notes":""},{"entity":{"id":"bbd594ac-365b-507f-954d-20a0df38312d","slug":"amn","display_name":"Human amnionless / AMN","entity_type_key":"protein"},"role":"subject","stoichiometry":null,"state_label":"","sequence_order":2,"notes":""},{"entity":{"id":"cca4766f-3ab0-57ea-8cb1-117af937c808","slug":"rat-mini-cubilin","display_name":"Rat N-terminal mini-cubilin construct","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":"Rat N-terminal mini-cubilin plus human AMN; HEK293T, MDCK, human RPTEC and HCT116","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"exposure","value_text":"Recombinant coexpression versus single transfection","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"limitations","value_text":"Mixed-species truncated construct lacks the complete CUB5-8 IF-binding region; surface expression is not direct B12 uptake.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"nutrient_topic","value_text":"Vitamin B12 research collection; topical membership is not evidence of a direct dietary effect.","comparator":null,"unit":null,"notes":"","entity":{"slug":"vitamin-b12","display_name":"Vitamin B12 (cobalamins)","entity_type_key":"chemical_species"}},{"dimension":"organism","value_text":"Rattus norvegicus construct; Homo sapiens AMN; human/canine host cells","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"plain_language","value_text":"Amnionless helped the experimental receptor reach the cell surface.","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"primary_references","value_text":"[udagawa-2018-glycosylation] Amnionless-mediated glycosylation is crucial for cell surface targeting of cubilin in renal and intestinal cells. (2018). https://pubmed.ncbi.nlm.nih.gov/29402915/ DOI: 10.1038/s41598-018-20731-4","comparator":null,"unit":null,"notes":"","entity":null},{"dimension":"tissue_or_cell_type","value_text":"Cultured renal and colorectal epithelial cells","comparator":null,"unit":null,"notes":"","entity":null}],"evidence":[{"id":"989a56dc-b6be-5fe4-8afe-5c6b5493cd53","evidence_kind":"source_excerpt","locator":"Lines 322-333","start_line":322,"end_line":333,"excerpt":"### b12-abs-amn-trafficking-rat-construct\nHuman AMN coexpression allowed rat mini-cubilin to reach the plasma membrane in HEK293T and other tested renal/intestinal cell models, whereas mini-cubilin alone remained intracellular.\nCondition category: normal\nnutrient_topic: Vitamin B12 research collection; topical membership is not evidence of a direct dietary effect.\nplain_language: Amnionless helped the experimental receptor reach the cell surface.\norganism: Rattus norvegicus construct; Homo sapiens AMN; human/canine host cells\ntissue_or_cell_type: Cultured renal and colorectal epithelial cells\nexperimental_model: Rat N-terminal mini-cubilin plus human AMN; HEK293T, MDCK, human RPTEC and HCT116\nlimitations: Mixed-species truncated construct lacks the complete CUB5-8 IF-binding region; surface expression is not direct B12 uptake.\nexposure: Recombinant coexpression versus single transfection\ncross_nutrient: false\n[udagawa-2018-glycosylation] Amnionless-mediated glycosylation is crucial for cell surface targeting of cubilin in renal and intestinal cells. (2018). https://pubmed.ncbi.nlm.nih.gov/29402915/ DOI: 10.1038/s41598-018-20731-4","model_system":"Rat N-terminal mini-cubilin plus human AMN; HEK293T, MDCK, human RPTEC and HCT116","directness":"author_interpretation","verification_status":"source_derived_draft","notes":"Exact curation-document quotation, not publisher quotation. Study references: [udagawa-2018-glycosylation] Amnionless-mediated glycosylation is crucial for cell surface targeting of cubilin in renal and intestinal cells. (2018). https://pubmed.ncbi.nlm.nih.gov/29402915/ DOI: 10.1038/s41598-018-20731-4","relationship":"supports","weight":1.0,"link_notes":"","source":{"id":"73145023-2982-5848-aff2-8d8875d6b9c5","stable_key":"import-7fc92b9e-9cbf-556e-8719-6b5244625250","title":"Vitamin B12: mechanisms, 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. 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