Homology modeling three-dimensional structure of AnxB1 and reducing its immunogenicity by sequence-deleted mutagenesis  

作　　者：YAN Hongli1, SONG Yunlong2, LIU Fan1, HE Yan1 & SUN Shuhan1 1. Department of Medical Genetics, Second Military Medical University, Shanghai 200433, China 2. Department of Medicinal Chemistry, Second Military Medical University, Shanghai 200433, China 

出　　处：《Science China(Life Sciences)》2004年第4期359-367,共9页中国科学（生命科学英文版）

基　　金：This work was supported by the National High Technology Research;Development Program of China(863 Program)(Grant No.101-060504);the National Natural Science Foundation of China(Grant No.30271167).

摘　　要：AnxB1, a novel annexin previously isolated from Cysticercus cellulose, shows high thrombi affinity and anticoagulant activity in vivo. In order to investigate the relationship between structure and biological function, a predicted three-dimensional (3D) model of AnxB1 was gen-erated by homology modeling. This model contains four homologous internal-domains and the Ca trace of domain I, II and IV shows high similarity. Based on the structure characterization, four sequence-deleted mutants were constructed and expressed as GST fusion proteins in E. coli. Two of the mutants, GST-M3 and GST-M4 reserved high anticoagulant activity (p<0.01 vs. GST). Furthermore, compared with the wild type GST-AnxB1, the immunogenicity of GST-M3 and GST-M4 was reduced significantly (p<0.01) and the molecular weight was lowered to 27 kD and 34 kD, respectively. These observations laid a solid foundation for further study on developing new thrombolytic agents with higher efficiency and lower side effect.AnxB1, a novel annexin previously isolated from Cysticercus cellulose, shows high thrombi affinity and anticoagulant activity in vivo. In order to investigate the relationship between structure and biological function, a predicted three-dimensional (3D) model of AnxB1 was gen-erated by homology modeling. This model contains four homologous internal-domains and the Ca trace of domain I, II and IV shows high similarity. Based on the structure characterization, four sequence-deleted mutants were constructed and expressed as GST fusion proteins in E. coli. Two of the mutants, GST-M3 and GST-M4 reserved high anticoagulant activity (p<0.01 vs. GST). Furthermore, compared with the wild type GST-AnxB1, the immunogenicity of GST-M3 and GST-M4 was reduced significantly (p<0.01) and the molecular weight was lowered to 27 kD and 34 kD, respectively. These observations laid a solid foundation for further study on developing new thrombolytic agents with higher efficiency and lower side effect.

关 键 词：annexin homology modeling sequence-deleted mutant anticoagulant immunogenicity. 

分 类 号：R346[医药卫生—基础医学]