%0 Journal Article
%T Simultaneous removal of T and B cell epitope in recombinant staphylokinase by structure-based mutagenesis of immuno-dominant Arg77 and Glu80 residues<br>Arg77和Glu80定点突变同时去除葡激酶中的T和B细胞抗原表位
%A Ruiguang Xu
%A Jintian He
%A Kai Ji
%A Xi Chen
%A Jianwei Liu
%A Ke Zhu
%A <br>徐瑞光
%A 贺进田
%A 贾锴
%A 陈希
%A 刘建伟
%A 朱科
%J 微生物学报
%D 2011
%I 
%X Objective]To reduce immunogenicity of recombined staphylokinase(r-yak),site-directed mutagenesis of Arg77 and Glu80 residue was performed to simultaneously remove T and B cell epitope in r-Sak molecule.Methods The solvent accessible surface areas of residues 77 and 80 in r-Sak were used to analyze rational design of Sak mutation.The Sak mutants were expressed in E coli DHSa.After purified by a 3-step chromatography,their fibrinolytic activities and immunological properties were analyzed.Results]Immunogenicity tests suggested that Sak induced a Th2-type immune response.Substitution of Glu80 with alanine or serine successfully reduced its solvent accessible surface area while simultaneously removing part of the T and B cell epitope.Changing Arg77 to glutamine,asparagine,or lysine removed only part of the T cell epitope.Of six dually substituted variants,Sak(R77Q/E80A)and Sak(R77Q/E80S)variants effectively eliminated part of the B and T cell epitopes,which markedly reduced their immunogenicity.
%K Keywords: staphylokinase
%K T cell epitope
%K B cell epitope
%K immunogenicity
%K protein engineering<br>葡激酶
%K T细胞抗原表位
%K B细胞抗原表位
%K 免疫原性
%K 蛋白质工程
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=89B49E12BCF515DCDF80DC28707FCFA6&yid=9377ED8094509821&vid=987EDA49D8A7A635&iid=94C357A881DFC066&sid=15F713CDE16A589C&eid=7D2B339649A57040&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=20