Maffei Benoit, Francetic Olivera, Subtil Agathe
Unité de Biologie Cellulaire de l'Infection Microbienne, Institut PasteurParis, France.
Centre National de la Recherche Scientifique UMR3691Paris, France.
Front Cell Infect Microbiol. 2017 May 31;7:221. doi: 10.3389/fcimb.2017.00221. eCollection 2017.
Bacteria have acquired multiple systems to expose proteins on their surface, release them in the extracellular environment or even inject them into a neighboring cell. Protein secretion has a high adaptive value and secreted proteins are implicated in many functions, which are often essential for bacterial fitness. Several secreted proteins or secretion machineries have been extensively studied as potential drug targets. It is therefore important to identify the secretion substrates, to understand how they are specifically recognized by the secretion machineries, and how transport through these machineries occurs. The purpose of this review is to provide an overview of the biochemical, genetic and imaging tools that have been developed to evaluate protein secretion in a qualitative or quantitative manner. After a brief overview of the different tools available, we will illustrate their advantages and limitations through a discussion of some of the current open questions related to protein secretion. We will start with the question of the identification of secreted proteins, which for many bacteria remains a critical initial step toward a better understanding of their interactions with the environment. We will then illustrate our toolbox by reporting how these tools have been applied to better understand how substrates are recognized by their cognate machinery, and how secretion proceeds. Finally, we will highlight recent approaches that aim at investigating secretion in real time, and in complex environments such as a tissue or an organism.
细菌已经获得了多种系统，用于将蛋白质暴露于其表面、释放到细胞外环境中，甚至将其注入相邻细胞。蛋白质分泌具有很高的适应性价值，分泌的蛋白质涉及许多功能，这些功能通常对细菌的适应性至关重要。几种分泌蛋白或分泌机制已被广泛研究作为潜在的药物靶点。因此，识别分泌底物、了解它们如何被分泌机制特异性识别以及如何通过这些机制进行转运非常重要。本综述的目的是概述已开发的用于定性或定量评估蛋白质分泌的生化、遗传和成像工具。在简要概述可用的不同工具之后，我们将通过讨论一些与蛋白质分泌相关的当前开放性问题来说明它们的优点和局限性。我们将从分泌蛋白的鉴定问题开始，这对许多细菌来说仍然是更好地理解它们与环境相互作用的关键第一步。然后，我们将通过报告这些工具如何被应用以更好地理解底物如何被其同源机制识别以及分泌如何进行来说明我们的工具箱。最后，我们将重点介绍旨在实时研究分泌以及在诸如组织或生物体等复杂环境中研究分泌的最新方法。
