Alternative split sites for fragment complementation,and glyphosate function as extra ligand and stabilizer for the AroA enzyme complexes  

作　　者：LI Yan SUN YiCheng YAN HaiQin WANG YiPing 

机构地区：[1]National Key Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China [2]Institute of Pathogen Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China

出　　处：《Chinese Science Bulletin》2011年第6期514-520,共7页

基　　金：supported by the National Transgenic Research and Industrialization Special Foundation of China (2009ZX08009057B and 2008ZX08009-003-002);the National Hi-Tech Research and Development Program of China (2007AA021307)

摘　　要：蛋白质体质分析能为蛋白质进化，蛋白质合拢和 macromolecular 汇编上的研究是有用的。AroA 是在向在微生物和植物的芳香的氨基酸的合成的小径的关键酶，并且是除草剂 glyphosate 的目标。我们的以前的学习证明从 Escherichia coli 的那功能的 AroA 酶能从蛋白质的二 220-amino 酸碎片被重新组成。在这研究，我们探索了 AroA 的这碎片互补。通过碎片互补的系统的研究，我们证明成功的碎片互补能在 vivo 被完成，当裂口地点在第二等的结构元素以内以及在在结构元素之间的环时。另外，第一次，我们提供证据那额外的 ligand 例如 glyphosate，能在 vivo 作为重新组成的建筑群的 stabilizer 工作。因此，我们的结果可以为蛋白质进化和复杂集会提供重要含意。Protein reconstitution analysis can be useful for studies on protein evolution, protein folding and macromolecular assembly. AroA is a key enzyme in the pathway toward the synthesis of aromatic amino acids in microorganisms and plants, and is the target of the herbicide glyphosate. Our previous study showed that functional AroA enzyme from Escherichia coli could be reconstituted from two -220-amino acid fragments of the protein. In this study, we explored this fragment complementation of AroA. Through a systematic study of fragment complementation, we show that successful fragment complementation can be achieved in vivo, when the split sites are within secondary structure elements as well as at loops between structure elements. In addition, we provide evidence, for the first time, that extra ligand, such as glyphosate, can function as a stabilizer of the reconstituted complexes in vivo Therefore, our results may provide important implications for protein evolution and complex assemblies.

关 键 词：复合稳定剂 酶复合物 草甘膦 互补 分裂 配体 蛋白质折叠 芳香氨基酸 

分 类 号：Q51[生物学—生物化学] TS275.4[轻工技术与工程—农产品加工及贮藏工程]