霍乱弧菌外切聚磷酸酶PPX的表达、纯化和初步晶体学研究  

Expression,Purification and Preliminary Crystallographic Analysis of Vibrio Cholerae Exopolyphosphatase PPX

作　　者：张皞 张琼林 Zhang Hao;Zhang Qionglin(College of Life Sciences,Nankai University,Tianjin 300071,China)

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《南开大学学报（自然科学版）》2024年第2期10-14,共5页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：国家自然科学基金面上项目(31570128)。

摘　　要：霍乱弧菌(Vibrio cholera)作为弧菌科的主要致病菌,是人类霍乱的病原体,它可以引起烈性肠道传染病,给人们的生命造成了巨大的威胁.多聚磷酸polyP广泛分布于细菌体内,在细菌的运动性、细胞膜的形成、严紧反应等过程中发挥关键作用,而外切聚磷酸酶PPX是水解polyP的主要蛋白,可直接影响细菌的致病性.本研究利用大肠杆菌原核表达的方法成功获得可溶性霍乱弧菌PPX蛋白,经过多种方式的纯化、结晶条件的筛选和优化得到蛋白晶体.通过X射线衍射实验收集到0.24 nm的衍射数据,使用HKL 2000软件对衍射数据进行了处理,这些数据为PPX晶体结构的解析奠定基础.As the main pathogen of Vibrio,is the pathogen of human cholera,which can cause severe intestinal infectious diseases and pose a great threat to people's lives.PolyP is widely distributed in bacteria,which plays a key role in the process of bacterial motility,cell membrane formation,and stringent reaction.Exopolyphosphatase PPX is the main protein hydrolyzing polyP,which can directly affect the pathogenicity of bacteria.In this study,the soluble PPX protein was successfully obtained by prokaryotic expression of Escherichia coli,and the protein crystals were obtained by screening and optimizing the crystallization conditions through various methods.Diffraction data of 0.24 nm were collected by X-ray diffraction experiments,and the diffraction data were processed using HKL2000.These data laid the foundation for the analysis of PPX crystal structure.

关 键 词：霍乱弧菌 外切聚磷酸酶 蛋白纯化 X射线晶体学 

分 类 号：Q71[生物学—分子生物学]