| 摘要: |
| 目的获取纯化的霍乱弧菌HutX蛋白晶体和衍射数据。方法通过基因克隆和蛋白表达得到HutX蛋白,通过镍柱亲和层析、Source Q阴离子交换柱层析和Superdex 200分子筛柱层析等纯化所得蛋白，并用蛋白质印迹法进行验证。将所获蛋白进行结晶条件筛选及悬滴优化，X线衍射分析所得晶体的结构。结果蛋白质印迹分析间接证明所得到的蛋白为HutX蛋白,并得到HutX蛋白的晶体及衍射数据。结论为进一步研究HutX的蛋白结构及其在霍乱弧菌亚铁血红素利用机制中发挥的作用提供了参考。 |
| 关键词: 霍乱弧菌 HutX蛋白 X线结晶学 血红素利用 |
| DOI：10.3724/SP.J.1008.2012.00969 |
| 投稿时间：2012-02-26修订日期：2012-06-21 |
| 基金项目:高等学校博士学科专项科研基金(20110131120045). |
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| Crystal structure of Vibrio cholerae HutX protein |
| LI Jun1,GONG Jing2,YANG Yong-ji1*,HUANG Yan3* |
(1. Department of Bio-physics, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China;
2. Center of Cancer Research, School of Medicine, Shandong University, Ji’nan 250012, Shandong, China
*Corresponding author.) |
| Abstract: |
| ObjectiveTo obtain purified Vibrio cholerae HutX and its diffraction data. MethodsProtein HutX was obtained by gene cloning and protein expression, purified by nickcl sepharose affinity chromatography, anion exchange chromatography (source Q), and molecular sieve chromatography (Superdex 200) , and identified by Western blotting analysis. Then the obtained protein was subjected to crystallization condition screening and hanging drop optimization. The obtained crystal structure was analyzed by X-ray diffraction method. ResultsWestern blotting analysis indirectly indicated that the obtained protein was HutX protein. Then the HutX crystal and its diffraction data were obtained in the present study. ConclusionThe findings of the present study pave a way for future research on the crystal structure and function of HutX protein and its role in heme utilization of Vibrio cholerae. |
| Key words: Vibrio cholerae HutX protein X-ray crystallography heme utilization |
