{"corpus_id":30309756,"paper_sha":"dc4b8b5e5d308a96974b9520845b604cf1060c8c","doi":"10.1074/jbc.M109.055061","arxiv_id":null,"pmid":19684016,"pmcid":"PMC2788834","mag_id":2085120816,"dblp_id":null,"acl_id":null,"title":"Single Chain Variable Fragment against Nicastrin Inhibits the γ-Secretase Activity*","year":2009,"publication_date":"2009-08-14","venue":"Journal of Biological Chemistry","journal":{"name":"The Journal of Biological Chemistry","pages":"27838 - 27847","volume":"284"},"journal_issn":null,"journal_title":null,"publication_types":["JournalArticle"],"pubmed_pub_types":["Journal Article","Research Support, Non-U.S. Gov't"],"s2_fields_of_study":["Biology","Medicine"],"reference_count":58,"citation_count":21,"influential_citation_count":0,"is_open_access":true,"arxiv_categories":null,"arxiv_license":null,"arxiv_journal_ref":null,"mesh_headings":[{"d":"Amino Acid Sequence","mj":false,"ui":"D000595"},{"d":"Amyloid Precursor Protein Secretases","mj":false,"qs":[{"q":"antagonists & inhibitors","mj":true,"ui":"Q000037"},{"q":"chemistry","mj":false,"ui":"Q000737"},{"q":"genetics","mj":false,"ui":"Q000235"},{"q":"immunology","mj":false,"ui":"Q000276"},{"q":"metabolism","mj":true,"ui":"Q000378"}],"ui":"D053829"},{"d":"Animals","mj":false,"ui":"D000818"},{"d":"Antibodies","mj":false,"qs":[{"q":"genetics","mj":false,"ui":"Q000235"},{"q":"immunology","mj":true,"ui":"Q000276"}],"ui":"D000906"},{"d":"Cell Line","mj":false,"ui":"D002460"},{"d":"Enzyme Inhibitors","mj":false,"qs":[{"q":"metabolism","mj":false,"ui":"Q000378"}],"ui":"D004791"},{"d":"Humans","mj":false,"ui":"D006801"},{"d":"Immunoglobulin Variable Region","mj":false,"qs":[{"q":"genetics","mj":false,"ui":"Q000235"},{"q":"metabolism","mj":true,"ui":"Q000378"}],"ui":"D007135"},{"d":"Indolizines","mj":false,"qs":[{"q":"metabolism","mj":false,"ui":"Q000378"}],"ui":"D007212"},{"d":"Membrane Glycoproteins","mj":false,"qs":[{"q":"chemistry","mj":true,"ui":"Q000737"},{"q":"genetics","mj":false,"ui":"Q000235"},{"q":"immunology","mj":true,"ui":"Q000276"}],"ui":"D008562"},{"d":"Molecular Sequence Data","mj":false,"ui":"D008969"},{"d":"Protein Binding","mj":false,"ui":"D011485"},{"d":"Protein Conformation","mj":false,"ui":"D011487"},{"d":"Protein Folding","mj":false,"ui":"D017510"},{"d":"Protein Structure, Tertiary","mj":false,"ui":"D017434"},{"d":"Sequence Alignment","mj":false,"ui":"D016415"},{"d":"Indolizidines","mj":false,"ui":"D054836"}],"chemicals":[{"n":"Amyloid Precursor Protein Secretases","ui":"D053829","reg":"EC 3.4.-"},{"n":"Antibodies","ui":"D000906","reg":"0"},{"n":"Enzyme Inhibitors","ui":"D004791","reg":"0"},{"n":"Immunoglobulin Variable Region","ui":"D007135","reg":"0"},{"n":"Indolizines","ui":"D007212","reg":"0"},{"n":"Membrane Glycoproteins","ui":"D008562","reg":"0"},{"n":"nicastrin protein","ui":"C413604","reg":"0"},{"n":"castanospermine","ui":"C037806","reg":"Q0I3184XM7"},{"n":"Indolizidines","ui":"D054836","reg":"0"}],"comments_corrections":null,"source_flags":5,"s2_open_access_pdf_url":"http://www.jbc.org/article/S0021925820383034/pdf","s2_open_access_landing_url":"https://www.semanticscholar.org/paper/dc4b8b5e5d308a96974b9520845b604cf1060c8c","s2_open_access_license":"CCBY","s2_open_access_status":"HYBRID","pmc_open_access_pdf_url":null,"pmc_open_access_landing_url":null,"pmc_open_access_license":null,"pmc_open_access_status":null,"unpaywall_open_access_pdf_url":null,"unpaywall_open_access_landing_url":null,"unpaywall_open_access_license":null,"unpaywall_open_access_status":null,"abstract":"γ-Secretase is a membrane protein complex that catalyzes intramembrane proteolysis of a variety of substrates including the amyloid β precursor protein of Alzheimer disease. Nicastrin (NCT), a single-pass membrane glycoprotein that harbors a large extracellular domain, is an essential component of the γ-secretase complex. Here we report that overexpression of a single chain variable fragment (scFv) against NCT as an intrabody suppressed the γ-secretase activity. Biochemical analyses revealed that the scFv disrupted the proper folding and the appropriate glycosyl maturation of the endogenous NCT, which are required for the stability of the γ-secretase complex and the intrinsic proteolytic activity, respectively, implicating the dual role of NCT in the γ-secretase complex. Our results also highlight the importance of the calnexin cycle in the functional maturation of the γ-secretase complex. The engineered intrabodies may serve as rationally designed, molecular targeting tools for the discovery of novel actions of the membrane proteins.","claims":[{"public_id":"cl_638c30a966c8dc5cdf98dd6980c11127","status":"active","text":"Overexpression of a single chain variable fragment against nicastrin as an intrabody suppressed γ-secretase activity.","confidence":0.98,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous (12632b8b5f)","roles":["extraction"],"url":"https://sah.borca.ai/u/12632b8b5f"}],"url":"https://sah.borca.ai/claims/cl_638c30a966c8dc5cdf98dd6980c11127"},{"public_id":"cl_bdba52ece1706c881062ba2d0c141337","status":"active","text":"Proper folding of nicastrin is required for γ-secretase complex stability, whereas glycosyl maturation is required for its intrinsic proteolytic activity.","confidence":0.93,"contributors":[{"id":1,"public_id":"12632b8b5f","public_label":"Anonymous 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