全面功能表征大肠杆菌脂质基因。

A Comprehensive Functional Characterization of Escherichia coli Lipid Genes.

机构信息

Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 2, 3584CM Utrecht, the Netherlands.

出版信息

Cell Rep. 2019 Apr 30;27(5):1597-1606.e2. doi: 10.1016/j.celrep.2019.04.018.

DOI:10.1016/j.celrep.2019.04.018

PMID:31042483

Abstract

Lipid membranes are the border between living cells and their environments. The membrane's lipid composition defines fluidity, thickness, and protein activity and is controlled by the intricate actions of lipid gene-encoded enzymes. However, a comprehensive analysis of each protein's contribution to the lipidome is lacking. Here, we present such a comprehensive and functional overview of lipid genes in Escherichia coli by individual overexpression or deletion of these genes. We developed a high-throughput lipidomic platform, combining growth analysis, one-step lipid extraction, rapid LC-MS, and bioinformatic analysis into one streamlined procedure. This allowed the processing of more than 300 samples per day and revealed interesting functions of known enzymes and distinct effects of individual proteins on the phospholipidome. Our data demonstrate the plasticity of the phospholipidome and unexpected relations between lipid classes and cell growth. Modeling of lipidomic responses to short-chain alcohols provides a rationale for targeted membrane engineering.

摘要

脂质膜是活细胞与其环境之间的边界。膜的脂质组成决定了流动性、厚度和蛋白质活性，并受到脂质基因编码酶的复杂作用的控制。然而，缺乏对每种蛋白质对脂质组贡献的全面分析。在这里，我们通过单独过表达或缺失这些基因，对大肠杆菌中的脂质基因进行了全面和功能的概述。我们开发了一种高通量脂质组学平台，将生长分析、一步法脂质提取、快速 LC-MS 和生物信息学分析结合到一个简化的流程中。这使得每天可以处理 300 多个样品，并揭示了已知酶的有趣功能以及单个蛋白质对磷脂组的不同影响。我们的数据表明了磷脂组的可变性，以及脂质类与细胞生长之间意想不到的关系。对短链醇的脂质组学反应进行建模，为有针对性的膜工程提供了依据。