BIOCHEMICAL CHARACTERIZATION OF BACTERIAL CYTOKINESIS

细菌细胞分裂的生化特征

• 批准号: 6739235
• 负责人: JOSEPH R MC CORMICK
• 金额: $ 3.1万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6739235
• 关键词: SDS polyacrylamide gel electrophoresis; Streptomyces; bacterial proteins; biochemistry; cell cycle; chromatography; crosslink; immunoprecipitation; mass spectrometry; membrane proteins; postdoctoral investigator; protein biosynthesis; protein degradation; protein structure function; protein transport; proteomics

DESCRIPTION (provided by applicant): Cell division will be biochemically analyzed in the filamentous prokaryote, Streptomyces coelicolor. This species is advantageous for this study because growth and viability do not require cell division, a collection of division null mutants already exists, and reagents vital to the success of this project have been prepared. Mounting evidence suggests that the division proteins function as a complex that is assembled in an ordered fashion; however direct biochemical support for the complex is absent. Many division proteins contain at east one membrane-spanning segment, making the biochemical characterization of the complex more challenging. Therefore, the time has come to apply membrane protein biochemical techniques, which will require training in the area. Specifically, I will identify the proteins in the Streptomyces cell division complex using three unique but complementary biochemical attacks: cross-linking, immune precipitation, and column Chromatography. These techniques have successfully defined other integral membrane protein machines. I also hope to identify novel components of the cell division complex using proteomics. The net result of this project will uniquely equip my laboratory to continue to make significant strides in the cell division field.

描述（由申请人提供）：细胞分裂将在丝状原核生物，链霉菌coelicolor中进行生化分析。该物种对本研究是有利的，因为生长和生存能力不需要细胞分裂，已经存在的分区无突变体的集合，并且已经准备了对该项目成功至关重要的试剂。越来越多的证据表明，分裂蛋白的作用是以有序的方式组装的复合物。但是，对复合物的直接生化支持是不存在的。许多分裂蛋白在East One跨膜段中包含，使该复合物的生化特征更具挑战性。因此，是时候应用膜蛋白生化技术了，这将需要该地区的培训。具体而言，我将使用三种独特但互补的生化攻击：交联，免疫沉淀和色谱柱色谱法鉴定链霉菌细胞分裂复合物中的蛋白质。这些技术已成功定义了其他积分膜蛋白机。我还希望使用蛋白质组学鉴定细胞分裂复合物的新成分。该项目的净结果将使我的实验室在细胞分裂领域继续取得长足的进步。