摘要
目的解析大肠杆菌T蛋白的四级结构。方法利用分段克隆法克隆表达了T蛋白及其两个独立结构域分支酸变位酶(CM,T/1-94)和预苯酸脱氢酶(PDH,T/93-373),再利用SDS-PAGE测得变性条件下T蛋白、CM和PDH的Mr分别为42×103,11×103和32×103,HPLC测得天然条件下CM和PDH的Mr分别为25×103和63×103,再用化学交联法分析聚合状态。结果显示T蛋白、独立结构域CM和PDH均为二聚体。结论说明大肠杆菌T蛋白二聚体是通过CM与CM、PDH与PDH相连的。
Abstract
OBJECTIVE To describe the tertiary structure of T-protein from Escherichia coli.METHODS T-protein,CM and PDH domains were cloned and expressed separately.The Mr determined by SDS-PAGE under denatured condition were 42×103,32×103 and 11×103 respectively for T-protein,PDH domain and CM domain,which were identical to the theoretical Mr.The calculated Mr of HPLC under the native condition were 63×103 and 25×103 respectively for PDH domain and CM domain.Chemical cross linking was employed to determine the polymerization status of T-protein,for its Mr was over-range in HPLC determination.RESULTS It is obvious that the Ms were doubled for PDH and CM domain under native condition.All of T-protein,CM and PDH are dimmers,Which was testified by both HPLC and chemical cross-linking experiments.CONCLUSION T-protein is a dimmer and it bonds together through adhesion of CM to CM and PDH to PDH domains.
关键词
大肠杆菌T蛋白 /
分支酸变位酶 /
预苯酸脱氢酶 /
聚合状态
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Key words
Escherichia coli T-protein /
Chorismate mutase /
Prephenate dehydrogenase /
Polymerization status
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王光柱;丁丁;孙红颖;陈枢青.
大肠杆菌T蛋白的聚合状态分析[J]. 中国药学杂志, 2006, 41(13): 1029-1032
WNG Gung-zhu;DING Ding;SUN Hong-ying;CHEN Shu-qing.
Polymerization Status Analysis of Escherichia coli T-protein [J]. Chinese Pharmaceutical Journal, 2006, 41(13): 1029-1032
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参考文献
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脚注
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基金
浙江省自然科学基金资助项目(302110);国家教育部回国人员启动基金(J20040141)
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