title: Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding.
abstract: Corin is a membrane-bound protease essential for activating natriuretic peptides and regulating blood pressure. Human corin has 19 predicted N-glycosylation sites in its extracellular domains. It has been shown that N-glycans are required for corin cell surface expression and zymogen activation. It remains unknown, however, how N-glycans at different sites may regulate corin biosynthesis and processing. In this study, we examined corin mutants, in which each of the 19 predicted N-glycosylation sites was mutated individually. By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. Similarly, N-glycosylation at Asn-231 protected corin from autocleavage in the frizzled-1 domain. Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. We also found that the location of the N-glycosylation site in the protease domain was not critical. N-Glycosylation at Asn-1022 may be switched to different sites to promote corin zymogen activation. Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin.
authors.affiliation: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195, the Department of Chemistry, Cleveland State University, Cleveland, Ohio 44115, and.
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.affiliation: the Cyrus Tang Hematology Center and Collaborative Innovation Center of Hematology, Soochow University, Suzhou 215123, China.
authors.forename: Tiantian
authors.initials: T
authors.lastname: Zhou
authors.affiliation: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors.forename: Jianhao
authors.initials: J
authors.lastname: Peng
authors.affiliation: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors.forename: Ping
authors.initials: P
authors.lastname: Xu
authors.affiliation: the Cyrus Tang Hematology Center and Collaborative Innovation Center of Hematology, Soochow University, Suzhou 215123, China.
authors.forename: Ningzheng
authors.initials: N
authors.lastname: Dong
authors.affiliation: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
authors.forename: Shenghan
authors.initials: S
authors.lastname: Chen
authors.affiliation: From the Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195, the Department of Chemistry, Cleveland State University, Cleveland, Ohio 44115, and the Cyrus Tang Hematology Center and Collaborative Innovation Center of Hematology, Soochow University, Suzhou 215123, China wuq@ccf.org.
authors.forename: Qingyu
authors.initials: Q
authors.lastname: Wu
mesh_terms: D000818:Animals; D009320:Atrial Natriuretic Factor / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D004792:Enzyme Precursors / Q000737:chemistry; D005434:Flow Cytometry; D006026:Glycoside Hydrolases / Q000737:chemistry; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D009154:Mutation; D032383:Myocytes, Cardiac / Q000166:cytology; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012697:Serine Endopeptidases / Q000737:chemistry*
chemical_list: D004792:Enzyme Precursors; D009320:Atrial Natriuretic Factor; D006026:Glycoside Hydrolases; C119812:CORIN protein, human; C485617:Corin protein, mouse; D012697:Serine Endopeptidases
keywords: Corin; Ectodomain Shedding; Intracellular Trafficking; Membrane Protease; N-Linked Glycosylation; Proteolytic Enzyme; Serine Protease; Shedding