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BIOCHEMICAL CHARACTERIZATION OF BACTERIAL CYTOKINESIS

细菌细胞分裂的生化特征

基本信息

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
项目类别：
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

项目摘要

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（由申请方提供）：将在丝状原核生物天蓝色链霉菌中对细胞分裂进行生化分析。该物种对于本研究是有利的，因为生长和活力不需要细胞分裂，已经存在分裂无效突变体的集合，并且已经制备了对本项目成功至关重要的试剂。越来越多的证据表明，分裂蛋白作为一个复杂的功能，组装在一个有序的方式，但直接的生物化学支持的复杂性是缺席。许多分裂蛋白含有至少一个跨膜片段，使得复合物的生物化学表征更具挑战性。因此，应用膜蛋白生物化学技术的时机已经到来，这将需要在该领域进行培训。具体来说，我将确定链霉菌细胞分裂复合物中的蛋白质，使用三种独特但互补的生化攻击：交联，免疫沉淀和柱层析。这些技术已经成功地定义了其他完整的膜蛋白机器。我还希望利用蛋白质组学来鉴定细胞分裂复合体的新成分。这个项目的最终结果将使我的实验室在细胞分裂领域继续取得重大进展。