牙龈卟啉单胞菌蛋白酶Tpr的表达、纯化和初步晶体学研究  

Expression,Purification,and Preliminary Crystallographic Analysis of Porphyromonas Gingivalis Protease Tpr

作　　者：张琼林 Mark Bartlam Zhang Qionglin Mark Bartlam(College of Life Science, 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）》2016年第5期14-19,共6页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：天津市应用基础研究计划项目(13JCYBJC20800);973计划前期研究专项(2014CB560709);国家自然科学基金面上项目(31570128)

摘　　要：牙龈卟啉单胞菌是一种能够引起牙周炎的口腔病原菌,能够分泌许多种蛋白酶.这些蛋白酶在获得营养、逃脱宿主免疫、粘连组织细胞和其它疾病进程中发挥了重要作用,被认为是牙周炎的重要致病因子.其中一种蛋白酶是巯基蛋白酶Tpr,它能够降解许多常见蛋白酶的底物,其活性受到营养状态和钙离子浓度的调节.本实验在大肠杆菌中表达得到Tpr蛋白,经多种方法纯化后筛选获得蛋白质晶体.通过X射线衍射实验收集到分辨率达到0.20 nm的衍射数据,并使用软件HKL2000进行了处理.这些数据为解析Tpr3维晶体结构奠定了基础.Porphyromonas gingivalis is an oral pathogen that causes human chronic periodontitis. It can express a variety of proteases, which play important roles in nutrient acquisition, host defense evasion, host tissues binding and other disease-related processes. Their strong activity is considered as an important virulence factor for periodontitis. One of the proteases is the thiol protease Tpr. It can degrade several gen- eral proteinase substrates and its activity is regulated by the nutrient conditions and the calcium concentra- tions. Here, we expressed Tpr in E.coli, purified Tpr using different methods and finally got well-diffract- ing protein crystals. X-ray diffraction data were collected to 0.20nm resolution and processed by the HKL2000 software. These data will make an important contribution to the structure determination of Tpr.

关 键 词：牙周炎 蛋白酶 TPR X射线衍射 晶体结构 

分 类 号：Q71[生物学—分子生物学]