结核分枝杆菌丝氨酸/苏氨酸蛋白激酶PknE胞外感受器结构域的晶体学初步研究

Preliminary Crystallographic Study of the Extracellular Sensor Domain of Mycobacterium Tuberculosis Serine/Threonine Protein Kinase PknE

结核分枝杆菌中的丝氨酸/苏氨酸蛋白激酶E(PknE)可以抑制宿主巨噬细胞的凋亡,从而使该病原菌得以在宿主中生存.PknE是由N端胞内激酶结构域与C端胞外感受器结构域组成,2个结构域通过单次跨膜螺旋连接.本实验在大肠杆菌表达系统中表达了PknE的胞外感受器结构域,将其纯化后对蛋白结晶条件进行了筛选,最终得到了形状较好的晶体.在上海同步辐射光源收集到分辨率达到0.175 nm的衍射数据,并对数据进行了初步的处理.这些实验和数据为PknE感受器结构域的结构解析奠定基础,并为基于结构的药物研发提供了重要信息.

The serine/threonine protein kinase E(PknE)in mycobacterium tuberculosis can inhibit apoptosis of macrophages to allow the pathogens surviving in the host.PknE consists of an N-terminal intracellular kinase domain and a C-terminal extracellular sensor domain,which are joined by a single transmembrane helix.In this experiment,the PknE extracellular sensor domain was cloned and expressed in the E.coli expression system.The protein crystallization conditions were screened after purification,and finally the crystal with a good shape was obtained.The diffraction data with the highest resolution of 0.175 nm was collected and processed in Shanghai Synchrotron Radiation Facility.All these experiments and data can lay the foundation for the structure determination of the PknE sensor domain which could be the useful information for structure-based drug discovery.