YidC-Structure, Function and Substrate Specificity

YidC-结构、功能和底物特异性

• 批准号: 1052033
• 负责人: Ross Dalbey
• 金额: $ 60万
• 依托单位: Ohio State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1052033&HistoricalAwards=false
• 关键词: YidC; Structure; Function; Substrate; Specificity

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.

智力优点这个项目的长期目标是了解蛋白质是如何插入生物膜的。 大多数细菌蛋白质通过Sec易位酶插入膜中，尽管一个子集通过YidC易位酶插入和折叠。该研究项目的目标是解决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YidC依赖于插入的膜蛋白的特征来确定YidC的底物特异性。 (2)阐明YidC在整合膜蛋白插入和折叠中的功能。(3)定义YidC单体的跨膜螺旋组织，并评估/扩展YidC二聚体模型。为了实现这些目标，将使用定点诱变、YidC耗竭菌株、蛋白质修饰、半胱氨酸交联和生物化学方法。 这些研究将为真核细胞中类似的插入途径提供线索，并为受体、通道、转运蛋白和ATP酶等膜蛋白的生物合成提供重要信息。这项研究的意义在于它靶向了一种重要而独特的蛋白质YidC，YidC是一种进化上保守的、参与膜蛋白生物合成的必需蛋白质。这个项目的结果将丰富我们对蛋白质插入和折叠的分子机器的理解，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训和教学本科生和研究生。 本科生培训的一个重要部分是激励学生做出发现，并允许他们测试假设。 该项目的学生成果将在会议上传播，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。 该实验室将招募和培训少数民族学生从事研究，并鼓励他们从事生命科学领域的研究生研究。 达尔比博士认为，必须在很小的时候就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。