title: Complement receptor activity of recombinant porcine CR1-like protein expressed in a eukaryotic system.
abstract: Primate complement receptor type 1 (CR1) protein, a single-chain transmembrane glycoprotein, plays an important role in immune adherence and clearing complement-opsonized immune complexes. Here, the mRNA of the porcine primate-like complement receptor (CR1-like) gene was analyzed, and two domain sequences with potential functions were cloned into the pwPICZalpha vector for expression in Pichia pastoris. The recombinant proteins were purified with both Protein Pure Ni-NTA resin and strong anion exchange resin. The activities of the purified recombinant proteins were evaluated by SDS-PAGE, western blotting, and complement receptor assays. The results indicated that two domains of the CR1-like protein, CCP36 and CCP811 with molecular weights of 29.8 kDa and 30 kDa, respectively, were successfully expressed in P. pastoris. These two recombinant proteins possess some of the functions of the primate CR1 protein. Using these two proteins coupled with an antibody blocking technique, we also showed that CR1-like is expressed on natural porcine erythrocytes.
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Yin
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Wei
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China.
authors.forename: Junbing
authors.initials: J
authors.lastname: Jiang
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China.
authors.forename: Kuohai
authors.initials: K
authors.lastname: Fan
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China.
authors.forename: Junxing
authors.initials: J
authors.lastname: Zhao
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China.
authors.forename: Na
authors.initials: N
authors.lastname: Sun
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Wang
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China.
authors.forename: Yaogui
authors.initials: Y
authors.lastname: Sun
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China.
authors.forename: Haili
authors.initials: H
authors.lastname: Ma
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhao
authors.affiliation: College of Animal Science and Veterinary Medicine, Shanxi Agriculture University, Taigu, 030801, Shanxi, People's Republic of China. livets@163.com.
authors.forename: Hongquan
authors.initials: H
authors.lastname: Li
mesh_terms: D000818:Animals; D000936:Antigen-Antibody Complex / Q000378:metabolism*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003167:Complement Activation; D004912:Erythrocytes / Q000378:metabolism*; D056890:Eukaryota; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D010843:Pichia / Q000235:genetics*; D011323:Primates / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics; D011951:Receptors, Complement / Q000235:genetics / Q000378:metabolism*; D040681:Structural Homology, Protein; D013552:Swine / Q000276:immunology*
chemical_list: D000936:Antigen-Antibody Complex; D012333:RNA, Messenger; D011951:Receptors, Complement
keywords: Biological activity; CR1-like; Erythrocyte; Immune adherence; Porcine; Yeast expression