DNA/RNA QUADRUPLEXES: STRUCTURE, STABILITY AND INTERACTIONS

DNA/RNA 四链体：结构、稳定性和相互作用

• 批准号: 7724191
• 负责人: RICHARD H SHAFER
• 金额: $ 0.61万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7724191
• 关键词: Affinity Chromatography; Amines; Apoptosis; Area; Biochemical; Biological Process; Biology; Cells; Chemotherapy-Oncologic Procedure; Complex; Computer Retrieval of Information on Scientific Projects Database; Engineering; Event; Funding; Goals; Grant; Institution; Label; Ligase; Light; Methods; Monitor; N-terminal; Pattern; Peptides; Play; Process; Proteins; Proteolysis; Proteolytic Processing; Proteomics; Regulation; Research; Research Personnel; Resources; Role; Sampling; Serum; Serum Proteins; Source; Staging; Stimulus; Structure; Surveys; United States National Institutes of Health; Work; base; cell type; chemotherapy; novel; tandem mass spectrometry

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. Proteolysis plays an important role in the regulation of diverse biological processes. One such process is apoptosis, in which approximately 300 proteins are known to undergo proteolytic processing as part of the biochemical events leading to programmed cell death. Unfortunately, current methods for monitoring proteolytic events in complex samples suffer from serious limitations. We are developing a novel method for global profiling of proteolysis in complex biochemical mixtures that is based on the use of an engineered peptide ligase to selectively label the alpha amines protein N-termini. The label permits affinity purification and enrichment of N-terminal peptides for subsequent sequencing by tandem mass spectrometry. This approach not only enables monitoring of newly formed N-termini as a result of proteolysis is cells or serum, but also decreases sample complexity for greater proteomic coverage. We are applying this method to profile the proteolysis that occurs during apoptosis in cells, and to profile changes in serum protein levels following apoptosis. Our goal is to survey the diversity of proteolysis patterns in apoptosis elicited by different stimuli and in different cell types, to identify new targets of proteolysis in apoptosis, and to explore how levels of serum proteins change in different stages of cancer and chemotherapy. This work will shed new light on the biology of apoptosis and how apoptosis can be best exploited in chemotherapy. The work will also establish a novel proteomic method that will find application in other areas of biology.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 蛋白水解酶在多种生物过程的调控中起着重要作用。其中一个这样的过程是细胞凋亡，在这个过程中，大约300个蛋白质被已知进行蛋白质分解处理，作为导致细胞程序性死亡的生化事件的一部分。不幸的是，目前监测复杂样品中蛋白质分解事件的方法受到严重限制。我们正在开发一种新的方法，用于对复杂的生化混合物中的蛋白质分解进行全局分析，该方法基于使用工程肽连接酶来选择性地标记阿尔法胺蛋白N末端。该标记允许对N-端肽进行亲和纯化和浓缩，以用于随后的串联质谱仪测序。这种方法不仅能够监测由于细胞或血清的蛋白质降解而形成的新形成的N-末端，而且还降低了样本的复杂性，以获得更大的蛋白质组覆盖率。我们正在应用这种方法来描述细胞在凋亡过程中发生的蛋白分解，以及在细胞凋亡后血清蛋白水平的变化。我们的目标是观察不同刺激和不同细胞类型诱导的细胞凋亡中蛋白降解模式的多样性，寻找新的细胞凋亡蛋白降解靶点，并探讨血清蛋白水平在癌症和化疗不同阶段的变化。这项工作将为细胞凋亡的生物学以及如何在化疗中最好地利用细胞凋亡提供新的线索。这项工作还将建立一种新的蛋白质组学方法，该方法将在生物学的其他领域得到应用。