title: Functional expression of the Fc-fused extracellular domains of group II membrane proteins.
abstract: The complicated delivery mechanism of group II membrane proteins makes it difficult to decide the fusion pattern of their extracellular domains (ECDs) with Fc moiety. In this study, we compared the expression of ECDs of three group II membrane proteins including CLEC-2, Dectin-1, and LOX-1 by fusion of Fc moiety. We found that the pattern of ECD-Fc fusion order produced the functionally active recombinant proteins while the pattern of Fc-ECD fusion order led to the altered glycosylation which abolished the binding of these proteins with their ligands. Meanwhile, our results indicated that the secretion of mouse Fc (mFc)-fused ECD of CLEC-2 was more efficient than that of rabbit Fc (rFc)-fused protein, while rFc moiety was more sensitive for detection compared with mFc moiety. Altogether, we provide a favorable fusion pattern of Fc moiety with the ECDs of group II transmembrane proteins.
authors.affiliation: Key Laboratory of Glycoconjugate Research, Ministry of Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, Shanghai, 200032, China.
authors.forename: Weicheng
authors.initials: W
authors.lastname: Wu
authors.forename: Jing
authors.initials: J
authors.lastname: Jin
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Liu
authors.forename: Yufei
authors.initials: Y
authors.lastname: Zhang
authors.forename: Meng
authors.initials: M
authors.lastname: Li
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Shao
authors.forename: Yifan
authors.initials: Y
authors.lastname: Qian
authors.forename: Dongmei
authors.initials: D
authors.lastname: Zhang
authors.forename: Haiyan
authors.initials: H
authors.lastname: Zhu
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Ruan
authors.forename: Jianhui
authors.initials: J
authors.lastname: Xie
authors.forename: Jianxin
authors.initials: J
authors.lastname: Gu
mesh_terms: D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism*; D037181:Lectins, C-Type / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D051127:Scavenger Receptors, Class E / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000099181:Dectin-1
chemical_list: D007141:Immunoglobulin Fc Fragments; D037181:Lectins, C-Type; D011994:Recombinant Proteins; D051127:Scavenger Receptors, Class E; D000099181:Dectin-1; C529518:CLEC-2 protein, mouse; C495138:Olr1 protein, mouse