YidC-Structure, Function and Substrate Specificity

这个项目的长期目标是了解蛋白质是如何插入生物膜的。大多数细菌蛋白是通过Sec转座酶插入到膜上的，尽管有一部分是通过YidC转座酶插入和折叠的。该研究项目的目标是研究YidC在膜蛋白三级结构生成中的新功能。该项目有三个重要目标：(1)通过鉴定使其依赖于YidC插入的膜蛋白的特征来确定YidC的底物特异性。(2)阐明YidC在整体膜蛋白插入和折叠中的功能。(3)明确YidC单体的跨膜螺旋结构，评价/扩展YidC二聚体模型。为了实现这些目标，将使用定点诱变、YidC耗尽菌株、蛋白质修饰、半胱氨酸交联和生化方法。这些研究将为真核细胞中类似的插入途径提供线索，并为膜蛋白（如受体、通道、转运体和atp酶）的生物发生提供重要信息。这项研究的意义在于，它针对的是一种重要而独特的蛋白YidC，它是一种进化上保守的、参与膜蛋白生物发生的必需蛋白。该项目的结果将丰富我们对蛋白质插入和折叠的分子机制的理解，并将有助于理解异常的膜蛋白折叠。更广泛的影响实验室积极培养和教学本科生和研究生。本科教育的一个重要组成部分是激励学生去发现并允许他们去检验假设。学生的研究成果将在会议上发表，并在顶级期刊上发表。膜蛋白插入的研究将在涉及分子生物学和生物化学方法的多学科研究环境中进行。该实验室将招收和培训少数民族学生从事研究，并鼓励他们在生命科学领域进行研究生研究。Dalbey博士认为，必须从小培养对生命科学和研究的兴趣。他将寻求与当地学校K-12的关系，向学生介绍科学发现的兴奋。

Intellectual MeritThe long-term objective of this project is to understand how proteins are inserted into biological membranes. The majority of bacterial proteins are inserted into the membrane by the Sec translocase, although a subset is inserted and folded by the YidC translocase. The goal of the research project is to address the novel function of YidC in the generation of the tertiary structure of membrane proteins. The project has three important aims: (1) Determine the substrate specificity of YidC by identifying the features of a membrane protein that make it YidC dependent for insertion. (2) Elucidate the function of YidC in the insertion and folding of integral membrane proteins. (3) Define the transmembrane helix organization of the YidC monomer, and evaluate/extend the YidC dimer model. To achieve these aims, site-directed mutagenesis, YidC depletion strains, protein modification, cysteine crosslinking, and biochemical methods will be used. These studies will provide clues to similar insertion pathway in eukaryotic cells and provide important information in the biogenesis of membrane proteins such as receptors, channels, transporters, and ATPases. This research is significant in that it targets an important and unique protein, YidC, an evolutionarily conserved and essential protein involved in membrane protein biogenesis. The results of this project will enrich our understanding of the molecular machines involved in the insertion and folding of proteins and will be useful in understanding aberrant membrane protein folding. Broader ImpactThe laboratory is active in training and teaching undergraduate and graduate students. An important part of the undergraduate training is to motivate students to make discoveries and allow them to test hypotheses. The student's results from this project will be disseminated at conferences and published in top-tiered journals. The research on membrane protein insertion will be performed in a multidisciplinary research environment involving molecular biology and biochemical approaches. The lab will recruit and train minority students in research and encourage them to pursue graduate research in the life sciences area. Dr. Dalbey believes that an interest in the life sciences and research must be cultivated at an early age. He will pursue relationships with local schools K-12 to introduce the students to the excitement of scientific discovery.