The requirement for PDIM in mycobacterial protein secretion

项目概要： 分枝杆菌蛋白质分泌的机制仍然难以捉摸。申请人的长期目标是 了解分枝杆菌蛋白分泌的分子机制。本申请的总体目标是- 方法是严格确定蛋白质分泌是否需要外层脂质PDIM（phthiocerol dimycerosate）， 是的。核心假设是PDIM促进分枝杆菌蛋白质在整个细胞中的最佳分泌。 分枝杆菌外膜（Mycobacterial outer membrane）。这项建议的基本理由是， 这项工作将可能为蛋白质分泌穿过包膜的难以捉摸的机制提供信息。 同时解决了本领域中关于PDIM在蛋白质分泌中的作用的未决问题。中央hypoth- 将通过以下具体目标测试：定义PDIM生物合成和运输的要求， 蛋白质分泌在这一目标下，遗传学与靶向和全球定量蛋白质组学的结合将 用于测量在存在或不存在PDIM产生的情况下蛋白分泌的变化，并定位 是的。成功完成拟议目标后的预期结果将是决定性的威慑- 确定分枝杆菌蛋白分泌对PDIM的需求。这一贡献将是巨大的- 因为它将直接告知分枝杆菌蛋白质分泌的难以捉摸的机制，并有可能识别 ESX-1基质，促进膜溶解，垂直推进场。此外，完成这一亲- 联合国儿童基金会将提供基本知识，对治疗和预防儿童疾病产生积极影响。 人类疾病，肺结核。这项拟议中的研究在概念上是创新的，因为它关注的是蛋白质 独立于一类分泌器的分泌。此外，该提案提供了一个创新的解释， 这可以统一文献中关于PDIM在蛋白质分泌中的作用的两个不同的研究。限定 外脂质PDIM与分泌的关系将通过提供新的见解而垂直地推进该领域 分枝杆菌蛋白质分泌的分子机制。

PROJECT SUMMARY: The mechanism of mycobacterial protein secretion remains elusive. The long-term goal of the applicant is to understand the molecular mechanisms of mycobacterial protein secretion. The overall objective of this applica- tion is to rigorously determine if the outer lipid, PDIM (phthiocerol dimycocerosate), is required for protein secre- tion. The central hypothesis is that PDIM promotes the optimal secretion of mycobacterial proteins across the MOM (mycobacterial outer membrane). The rationale underlying this proposal is that the successful completion of the proposed work will potentially inform the elusive mechanisms of protein secretion across the envelope while resolving an open question in the field regarding the role of PDIM in protein secretion. The central hypoth- esis will be tested by following this specific aim: Define the requirement for PDIM biosynthesis and transport in protein secretion. Under this aim, a combination of genetics with targeted and global quantitative proteomics will be used to measure changes in protein secretion in the presence or absence of PDIM production and localiza- tion. The expected outcome following the successful completion of the proposed aim will be the definitive deter- mination of the requirement of PDIM in mycobacterial protein secretion. This contribution will be significant be- cause it will directly inform the elusive mechanism of mycobacterial protein secretion and potentially identify the ESX-1 substrates that promote membrane lysis, advancing the field vertically. Moreover, completion of this pro- posal will contribute fundamental knowledge that could positively impact the treatment and prevention of the human disease, tuberculosis. The proposed research is conceptually innovative because it focuses on protein secretion independent of a class secretory apparatus. Moreover, the proposal provides an innovative explanation which could unify two disparate studies in the literature regarding the role of PDIM in protein secretion. Defining the relationship of the outer lipid PDIM with secretion will advance the field vertically by providing new insight into the molecular mechanisms of mycobacterial protein secretion.