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口腔致病菌变形链球菌Smu.776蛋白的制备、结晶及初级晶体学分析
引用本文:王开拓,李兰芬,张晓嫣,梁宇和,魏世成,苏晓东.口腔致病菌变形链球菌Smu.776蛋白的制备、结晶及初级晶体学分析[J].生物化学与生物物理进展,2007,34(2):176-179.
作者姓名:王开拓  李兰芬  张晓嫣  梁宇和  魏世成  苏晓东
作者单位:1. 蛋白质工程和植物基因工程国家重点实验室,北京大学生命科学学院,北京,100871
2. 北京大学口腔医学院,北京,100081
基金项目:国家自然科学基金;面向21世纪教育振兴行动计划(985计划);北京大学校科研和教改项目;教育部霍英东教育基金
摘    要:变形链球菌smu.776基因编码一段含有385个氨基酸的蛋白质,其功能可能为依赖于腺苷甲硫氨酸的甲基转移酶.smu.776的DNA片段被克隆到表达载体pET28a后,在大肠杆菌BL21(DE3)菌株中过量表达得到很好的产量.产物Smu.776蛋白通过Ni2 亲和柱和分子筛柱层析两步得到纯化.采用悬滴气象扩散法得到了Smu.776蛋白的晶体.X射线衍射分辨率达到2.0!,晶体属单斜空间群C2,晶格参数为a=168.47",b=50.66#,c=53.96$,β=104.22°.每个最小不对称单元内含有一个蛋白质分子,溶剂含量为51.3%.

关 键 词:变形链球菌  龋齿  蛋白晶体学
收稿时间:2006/6/30 0:00:00
修稿时间:8/2/2006 12:00:00 AM

Preparation, Crystallization and Preliminary X-ray Crystallographic Analysis of Smu.776 From Caries Pathogen Streptococcus mutans
WANG Kai-Tuo,LI Lan-Fen,ZHANG Xiao-Yan,LIANG Yu-He,WEI Shi-Cheng and SU Xiao-Dong.Preparation, Crystallization and Preliminary X-ray Crystallographic Analysis of Smu.776 From Caries Pathogen Streptococcus mutans[J].Progress In Biochemistry and Biophysics,2007,34(2):176-179.
Authors:WANG Kai-Tuo  LI Lan-Fen  ZHANG Xiao-Yan  LIANG Yu-He  WEI Shi-Cheng and SU Xiao-Dong
Institution:National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China;National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China;National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China;National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China;National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China
Abstract:The gene smu. 776 encodes a possible S-adenosylmethionine-dependent methyltransferase of 385 residues in Streptococcus mutans, a primary pathogen for human dental caries. The DNA fragment of smu.776 was cloned into pET28a and expressed in good amount from the E. coli strain BL21 (DE3). Smu.776 protein was purified to homogeneity in a two-step procedure ofNi2+ chelating and size exclusion chromatography. Crystals were obtained by hanging-drop vapor diffusion method and diffracted to 2.0 (A) resolution.The crystal belongs to orthorhombic space group C2 with cell dimension of a=168.47 (A), b= 50.66 (A), c=53.96 (A), β=104.22°. The asymmetric unit is expected to contain one molecule with solvent content of 51.3%.
Keywords:Smu  776  Streptococcus mutans  dental caries  Smu  776  protein crystallography
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