Silent mutations affect in vivo protein folding in Escherichia coli
Silent mutations affect in vivo protein folding in Escherichia coli
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DOI：
10.1016/s0006-291x(02)00226-7
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发表时间：
2002-04-26
影响因子：
3.1
通讯作者：
Deana, A
中科院分区：
文献类型：
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作者：
Cortazzo, P;Cerve単ansky, C;Deana, A
为了研究体内蛋白质折叠的分子机制以及翻译动力学在特定折叠途径中的作用，我们对EgFABP1（棘球绦虫脂肪酸结合蛋白1）基因进行了密码子替换，将5个次要密码子替换为它们的同质主密码子。改变的区域对应于两个短α螺旋之间的旋转。当EgFABP1在大肠杆菌中表达时，其中一个沉默突变显著降低了该蛋白的溶解度。该蛋白的表达还引起了一个旨在检测错误折叠蛋白的报告基因的强烈激活，这表明该翻转区域似乎具有特殊的翻译动力学要求，以确保蛋白质的正确折叠。我们的研究结果强调了密码子使用在体内蛋白质折叠中的重要性。(C) 2002 Elsevier Science （USA）。版权所有。
As an approach to investigate the molecular mechanism of in vivo protein folding and the role of translation kinetics on specific folding pathways, we made codon substitutions in the EgFABP1 (Echinococcus granulosus fatty acid binding protein1) gene that replaced five minor codons with their synonymous major ones. The altered region corresponds to a turn between two short alpha helices. One of the silent mutations of EgFABP1 markedly decreased the solubility of the protein when expressed in Escherichia coli. Expression of this protein also caused strong activation of a reporter gene designed to detect misfolded proteins, suggesting that the turn region seems to have special translation kinetic requirements that ensure proper folding of the protein. Our results highlight the importance of codon usage in the in vivo protein folding. (C) 2002 Elsevier Science (USA). All rights reserved.