TARGETED PROTEOMIC STUDY OF THE CYCLIN-CDK MODULE

CYCLIN-CDK 模块的靶向蛋白质组学研究

• 批准号: 8169131
• 负责人: Brian T Chait
• 金额: $ 2.33万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169131
• 关键词: Adaptor Signaling Protein; Binding; Biological; Cell Cycle; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Cyclin-Dependent Kinases; Cyclins; Data; Degradation Pathway; Detection; Funding; Grant; Institution; Mass Spectrum Analysis; Phosphorylation; Proteins; Proteome; Proteomics; Reporting; Research; Research Personnel; Resources; Route; Saccharomyces cerevisiae; Saccharomycetales; Source; Specificity; Ubiquitination; United States National Institutes of Health; Work; base; inhibitor/antagonist; research study; time use

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The cell division cycle of the budding yeast Saccharomyces cerevisiae is mainly driven by one Cdk (Cyclin-Dependent Kinase), which becomes active when bound to one of nine cyclin subunits. The majority of the specific associates and substrates for these cyclin-Cdk complexes remain elusive, though their elucidation is essential for a full understanding of the cell cycle. Here we report the results of a targeted proteomics study that identified numerous proteins associated with particular cyclin-Cdk complexes. These included phosphorylation substrates, proteins involved in the ubiquitination-degradation pathway, adaptor proteins and inhibitors. We investigated and confirmed the specificity and biological relevance of some of these interactions. We demonstrate that this approach for studying protein associations within a biological module allowed the detection of many new associations. Our data includes many associations that were missed in previous proteome-wide studies, and shows that even transient and dynamic interactions can be detected by mass spectrometry-based targeted proteomic approaches. This work was presented in Archambault V, Chang EJ, Drapkin BJ, Cross FR, Chait BT, Rout MP, Targeted proteomic study of the cyclin-Cdk module Mol Cell. 2004, 14, 699-711. We are currently revisiting this experiment under conditions that have been highly optimized over the past several years, as well as using time-resolved proteomics and optimized mass spectrometric readout. Under these new conditions, we hope to isolate & identify a large number of new substrates of Cdk.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 芽殖酵母酿酒酵母的细胞分裂周期主要由一种Cdk（细胞周期蛋白依赖性激酶）驱动，当与九种细胞周期蛋白亚基之一结合时，Cdk变得活跃。大多数特定的联营公司和这些细胞周期蛋白-Cdk复合物的基板仍然难以捉摸，虽然他们的澄清是必不可少的细胞周期的全面了解。 在这里，我们报告的结果，有针对性的蛋白质组学研究，确定了许多蛋白质与特定的细胞周期蛋白-Cdk复合物。 这些包括磷酸化底物，参与泛素化降解途径的蛋白质，衔接蛋白和抑制剂。我们研究并证实了其中一些相互作用的特异性和生物学相关性。 我们证明，这种方法用于研究蛋白质协会内的生物模块允许检测到许多新的协会。 我们的数据包括许多在以前的蛋白质组研究中遗漏的关联，并表明即使是瞬时和动态的相互作用也可以通过基于质谱的靶向蛋白质组学方法检测到。这项工作发表在Archambault V，Chang EJ，Drapkin BJ，Cross FR，Chait BT，Rout MP，Targeted Proteomic study of the cyclin-Cdk module Mol Cell. 2004，14，699-711。我们目前正在重新审视这个实验，在过去几年中已经高度优化的条件下，以及使用时间分辨蛋白质组学和优化的质谱读数。在这些新的条件下，我们希望分离和鉴定大量的Cdk的新底物。