| 龋齿致病菌变形链球菌蛋白 Smu.260 的 制备、结晶及初级晶体学分析 |
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| 引用本文： | 张晓嫣,米 伟,周艳峰,刘翔宇,李兰芬,梁宇和,魏世成,苏晓东.龋齿致病菌变形链球菌蛋白 Smu.260 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,2005,32(3):217-220. |
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| 作者姓名： | 张晓嫣 米 伟 周艳峰 刘翔宇 李兰芬 梁宇和 魏世成 苏晓东 |
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| 作者单位： | 1. 北京大学生命科学学院,蛋白质工程和植物基因工程国家实验室,北京,100871;北京大学生命科学学院生物化学与分子生物学系,北京,100871
2. 北京大学口腔医学院,北京,100081 |
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| 基金项目： | 国家高技术“863”计划资助项目（2002BA711A13）及北京大学985和211工程项目资助. 苏晓东教授为国家自然科学基金杰出青年基金获得者(30325012). |
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| 摘 要： | 变形链球菌 (Streptococcus mutans) 是最主要的龋齿致病菌，其基因 Smu.260 编码一个约 23 ku (200 个氨基酸 ) 的蛋白质. Smu.260的 DNA 片段被克隆到表达载体 pET28a 后在大肠杆菌 BL21(DE3) 菌株中表达得到很好的产量. 产物 Smu.260 蛋白通过 Ni2+亲和柱和分子筛两步法纯化，并发现纯化后的蛋白以两种形式存在，二聚体 (约46 ku) 和四聚体，前者呈亮黄色，后者无色. 采用悬滴气象扩散法得到了二聚体形式的晶体. 晶体的 X 射线衍射分辨率达到 2.3埃，晶体属正交空间群 P212121，晶格参数为a=89.88埃, b=90.91埃, c=105.17埃. 晶胞不对称单元内估计含有一个二聚体，溶剂含量为 53％ .
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| 关 键 词： | 变形链球菌，龋齿， Smu.260 ，蛋白质晶体学 |
Protein Preparation, Crystallization and Preliminary X-ray Crystallographic Analysis of Smu.260 From Streptococcus mutans--a Cariogenic Dental Pathogen |
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| ZHANG Xiao-Yan,MI Wei,ZHOU Yan-Feng,LIU Xiang-Yu,LI Lan-Fen,LIANG Yu-He,WEI Shi-Cheng and SU Xiao-Dong.Protein Preparation, Crystallization and Preliminary X-ray Crystallographic Analysis of Smu.260 From Streptococcus mutans--a Cariogenic Dental Pathogen[J].Progress In Biochemistry and Biophysics,2005,32(3):217-220. |
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| Authors: | ZHANG Xiao-Yan MI Wei ZHOU Yan-Feng LIU Xiang-Yu LI Lan-Fen LIANG Yu-He WEI Shi-Cheng and SU Xiao-Dong |
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| Institution: | 1)National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing 100871, China;2)Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, Beijing 100871, China;1)National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing 100871, China;2)Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, Beijing 100871, China;1)National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing 100871, China;2)Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, Beijing 100871, China;1)National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing 100871, China;2)Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, Beijing 100871, China;1)National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing 100871, China;2)Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, Beijing 100871, China;1)National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing 100871, China;2)Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, Beijing 100871, China;Peking University School of Stomatology, Beijing 100081, China;1)National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing 100871, China;2)Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, Beijing 100871, China |
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| Abstract: | Smu.260 encodes a putative protein of 200 residues in Streptococcus mutans, a primary pathogen for human dental caries. Smu.260 was cloned into expression vector pET28a and expressed in good amount from the E.coli strain BL21 (DE3). Smu.260 protein was purified to homogeneity in a two-step procedure of Ni2+ chelating and size exclusion chromatography. The purified protein exists in two forms, a dimer form about 46 ku with yellow color and a tetramer form without apparent color. Crystals were obtained from the dimer protein by hanging-drop vapor-diffusion method. The crystals diffracted to about 2.3 Amstrong resolution and belong to orthorhombic space group P212121 with cell dimensions of a=89.88 Amstrong, b=90.91 Amstrong, c=105.17 Amstrong. The asymmetric unit is expected to contain two dimers with solvent content of 53%. |
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| Keywords: | Streptococcus mutans dental caries Smu 260 protein crystallography |
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