The integration of tail anchored membrane proteins by the twin-arginine translocase

双精氨酸转位酶对尾锚定膜蛋白的整合

• 批准号: BB/S005307/1
• 负责人: Tracy Palmer
• 金额: $ 59.3万
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FS005307%2F1
• 关键词: integration; tail; anchored; membrane; proteins

All bacteria, whether they are 'friendly' or otherwise, have one thing in common. In order to colonize their niches they need to communicate with the outside world. They achieve this by secreting protein molecules that allow them firstly to detect and then manipulate their environment. Bacteria are surrounded by one or more membranes and a rigid wall, which together form a protective barrier. Secreting proteins into the environment requires that these molecules are able to pass through the membrane barrier. In order to achieve this, bacteria have transporters located in the membrane that allow the passage of proteins to the outside. Understanding how these protein transporters work is critical if we wish to control this process to prevent disease, or engineer microbes to decontaminate toxic environments.We study a protein transporter called the Tat system, that is conserved in almost all bacteria and in plant chloroplasts, and we use E. coli as a convenient model system in which to study these processes. The Tat system plays a very important role in the physiology of many different bacteria and it is essential for photosynthesis in plants. Proteins are made up of long, linear chains of amino acids which fold up after they are made. Proteins are only functional once they have folded into their final 3-dimensional structure. Proteins that are secreted are functional outside the bacterial cell. The Tat system is unusual because unlike most other protein transporters it only transports proteins after they have already folded. Proteins that are destined to be secreted by the Tat system have a special signature sequence of amino acids, termed a 'twin arginine signal' at their start. This signal targets the protein to the Tat machinery that is embedded in the membrane, and facilitates its secretion. The Tat machinery itself is made up of three components - TatA, TatB and TatC. The TatA, TatB and TatC components initially form a 1:1:1 complex with each other and this complex is responsible for recognizing each of the different proteins that are targeted for secretion by Tat system, by interacting with the twin arginine signal. After the signal has been bound by TatABC this triggers additional copies of TatA component to assemble into a ring-like structure, which can then allow transport of the protein. After the protein has been transported the TatA ring disassembles ready for another round of secretion.Some of the Tat substrate proteins are unusual because they contain a membrane interaction domain at their C-terminus that anchors them into the membrane. In E. coli these membrane-anchored Tat substrates are essential for the bacterium to adapt to different growth conditions, and to survive during infection of mammals. We are interested in understanding how the Tat pathway is able to integrate these proteins into the membrane. We have isolated mutants in the Tat system that cannot integrate these proteins, instead secreting them completely across the membrane. We want to understand why these mutants are defective in integrating the Tat substrate proteins. This work will help us to understand how the Tat machinery recognises membrane proteins, and may also help us to understand the mechanism by which the Tat machinery disassembles once transport is complete.

所有的细菌，不管它们是“友好的”还是其他的，都有一个共同点。为了开拓他们的利基市场，他们需要与外界沟通。它们通过分泌蛋白质分子来实现这一点，这些蛋白质分子使它们能够首先检测并操纵它们的环境。细菌被一个或多个膜和刚性壁包围，它们共同形成保护屏障。将蛋白质分泌到环境中需要这些分子能够穿过膜屏障。为了实现这一点，细菌具有位于膜中的转运蛋白，允许蛋白质通过到外部。如果我们希望控制这一过程以预防疾病，或设计微生物以净化有毒环境，那么了解这些蛋白质转运蛋白的工作方式至关重要。我们研究了一种称为达特系统的蛋白质转运蛋白，该系统在几乎所有细菌和植物叶绿体中都是保守的。大肠杆菌作为一个方便的模型系统，在其中研究这些过程。达特系统在许多不同细菌的生理学中起着非常重要的作用，并且它对于植物的光合作用是必不可少的。蛋白质是由长的线性氨基酸链组成的，这些氨基酸链在形成后折叠起来。蛋白质只有在折叠成最终的三维结构后才有功能。分泌的蛋白质在细菌细胞外起作用。达特系统是不寻常的，因为不像大多数其他蛋白质转运蛋白，它只运输蛋白质后，他们已经折叠。注定由达特系统分泌的蛋白质具有特殊的氨基酸特征序列，在其起始处称为“双精氨酸信号”。该信号将蛋白质靶向嵌入膜中的达特机制，并促进其分泌。达特机器本身由三个部件组成- TatA、TatB和TatC。TatA、TatB和TatC组分最初彼此形成1：1：1复合物，并且该复合物负责通过与双精氨酸信号相互作用来识别被达特系统靶向分泌的不同蛋白质中的每一种。在信号被TatABC结合后，这触发了TatA组分的额外拷贝组装成环状结构，然后可以允许蛋白质的运输。在蛋白质被转运后，TatA环被分解，准备进行另一轮分泌。一些达特底物蛋白是不寻常的，因为它们在C末端含有一个膜相互作用结构域，将它们锚定在膜上。在大肠这些膜锚定的达特底物对于细菌适应不同的生长条件以及在感染哺乳动物期间存活是必需的。我们有兴趣了解达特途径是如何能够整合这些蛋白质进入膜。我们已经在达特系统中分离出了突变体，它们不能整合这些蛋白质，而是完全分泌它们穿过膜。我们想了解为什么这些突变体在整合达特底物蛋白方面有缺陷。这项工作将帮助我们了解达特机器如何识别膜蛋白，也可能帮助我们了解达特机器在运输完成后分解的机制。

项目成果

The Carbapenemase BKC-1 from Klebsiella pneumoniae Is Adapted for Translocation by Both the Tat and Sec Translocons.

• DOI: 10.1128/mbio.01302-21
• 发表时间: 2021-06-29
• 期刊: mBio
• 影响因子: 6.4
• 作者: Bharathwaj M;Webb CT;Vadlamani G;Stubenrauch CJ;Palmer T;Lithgow T
• 通讯作者: Lithgow T

Ferric Citrate Regulator FecR Is Translocated across the Bacterial Inner Membrane via a Unique Twin-Arginine Transport-Dependent Mechanism.

柠檬酸铁调节剂 FecR 通过独特的双精氨酸运输依赖机制跨细菌内膜转运。

• DOI: 10.1128/jb.00541-19
• 发表时间: 2020
• 期刊: Journal of bacteriology
• 影响因子: 3.2
• 作者: Passmore IJ