Alanine scanning mutagenesis identifies an asparagine-arginine-lysine triad essential to assembly of the shell of the Pdu microcompartment.
Alanine scanning mutagenesis identifies an asparagine-arginine-lysine triad essential to assembly of the shell of the Pdu microcompartment.
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DOI：
10.1016/j.jmb.2014.04.012
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发表时间：
2014-06-12
影响因子：
5.6
通讯作者：
Bobik, Thomas A.
中科院分区：
文献类型：
--
作者：
Sinha, Sharmistha;Cheng, Shouqiang;Sung, Yea Won;McNamara, Dan E.;Sawaya, Michael R.;Yeates, Todd O.;Bobik, Thomas A.
细菌微区室（MCP）是已知的最简单的细胞器。它们的作用是通过将几种相关酶限制在称为外壳的全蛋白质包膜内来增强代谢途径。在这项研究中，我们调查的因素，支配MCP大会进行扫描诱变的表面残基的PduA，一个主要的壳蛋白的MCP用于1，2-丙二醇降解。对20个突变体的生化、遗传和结构分析使我们能够确定PduA K26、N29和R79是稳定1，2-丙二醇MCP的壳的关键残基。此外，我们确定了两个PduA突变体（K37 A和K55 A），最有可能通过改变其蛋白质壳的渗透性来损害MCP功能。这是第一次研究在微区室系统中壳蛋白结构突变的表型效应。这里报道的研究结果可能适用于工程蛋白质容器，具有改善的生物技术应用的稳定性。
Bacterial microcompartments (MCPs) are the simplest organelles known. They function to enhance metabolic pathways by confining several related enzymes inside an all protein envelope called the shell. In this study, we investigated the factors that govern MCP assembly by performing scanning mutagenesis on the surface residues of PduA, a major shell protein of the MCP used for 1,2-propanediol degradation. Biochemical, genetic and structural analysis of 20 mutants allowed us to determine that PduA K26, N29 and R79 are crucial residues that stabilize the shell of the 1,2-propanediol MCP. In addition, we identify two PduA mutants (K37A and K55A) that impair MCP function most likely by altering the permeability of its protein shell. These are the first studies to examine the phenotypic effects of shell protein structural mutations in a microcompartment system. The findings reported here may be applicable to engineering protein containers with improved stability for biotechnology applications.
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影响因子：
8
作者：
Dryden, Kelly A.;Crowley, Christopher S.;Yeager, Mark
通讯作者：
Yeager, Mark
影响因子：
3.2
作者：
Johnson, CLV;Pechonick, E;Bobik, TA
通讯作者：
Bobik, TA
影响因子：
3.2
作者：
BERKOWIT.D;HUSHON, JM;AMES, BN
通讯作者：
AMES, BN
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
3.2
作者：
Havemann, GD;Sampson, EM;Bobik, TA
通讯作者：
Bobik, TA