Mechanism and Dynamics of the YidC Insertase in Membrane Protein Insertion

该项目研究了名为YidC的生物机器如何将蛋白质插入并折叠到膜双层中。蛋白质插入膜是生物学中最关键的过程之一，了解蛋白质跨膜运输或整合到膜中的机制对于了解细胞和生物体的结构，功能和健康具有巨大的潜在影响。该项目将产生尚未发现的关于YidC功能的基本信息，这是理解生物膜如何组装，保持其身份并支持信号传导和运输等许多功能的关键。由此产生的关于膜蛋白生物发生的新知识也可能影响未来与人类健康相关的研究，因为膜蛋白占已知药物靶点的50%以上。该项目还将提供机会，在涉及分子生物学、生物化学和生物物理方法的多学科研究环境中培训学生。 这项研究的目的是确定YidC如何将蛋白质插入细胞膜。YidC作为膜插入酶是新颖的，因为它含有对其插入酶和伴侣蛋白功能至关重要的亲水性空腔。YidC可以单独发挥作用，也可以与Sec机制合作插入蛋白质。该项目利用荧光光谱，电子顺磁共振自旋标记和单分子方法来检查膜插入事件的动力学和结构关系。该研究的具体目的是确定YidC在膜蛋白插入中的作用机制和动力学，阐明YidC/Sec机械复合物中的相互作用，并确定决定E.杆菌该项目将推动细菌膜蛋白插入领域的发展，并将影响对人类细胞中类似途径的理解。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This project examines how the bio-machine named YidC inserts and folds proteins into the membrane bilayer. Protein insertion into membranes is one of the most critical processes in biology, and understanding the mechanisms by which proteins are transported across or integrated into membranes has enormous potential impact for understanding cellular and organismal structure, function and health. The project will yield, as yet undiscovered, fundamental information on the function of YidC, which is key to understanding how biological membranes are assembled, maintain their identity, and support many functions such as signaling and transport. The resulting new knowledge on biogenesis of membrane proteins could also impact human health-related research in the future since membrane proteins make up more than 50% of the known drug targets. The project will also provide opportunities for training students in a multidisciplinary research environment involving molecular biology, biochemical and biophysical approaches. The goal of the research is to determine how YidC functions to insert proteins into the cell membrane. YidC is novel as a membrane insertase as it contains a hydrophilic cavity that is crucial for its insertase and chaperone function. YidC can function on its own or cooperate with the Sec machinery to insert proteins. The project utilizes fluorescence spectroscopy, electron paramagnetic resonance spin labeling, and single molecule approaches to examine the dynamics and structural relationships of membrane insertion events. The research specifically aims to determine the mechanism of action and dynamics of YidC in membrane protein insertion, elucidate the interactions in the YidC/Sec machinery complex, and identify features of membrane protein substrates that determine selection of YidC and Sec translocase pathways in E. coli. The project will advance the field of membrane protein insertion in bacteria and will impact understanding of similar pathways in human cells.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.