Profiling of Protein Modifications by Mass Spectrometry

描述：(申请人提供)：细胞信号通路有多种类型的调节，从转录控制到蛋白质降解的调节。这些过程通常由相关蛋白质的酶修饰来调节，因此蛋白质的修饰是大多数信号转导途径中不可或缺的一部分。在这个项目中，我们计划开发检测蛋白质组内修饰的方法。我们最初的重点将是检测癌症中蛋白质的酪氨酸磷酸化。我们计划开发丰富修饰蛋白质的细胞提取物的方法，直接耦合到基于质谱学的检测和鉴定这些蛋白质的方法。我们还计划通过将这些“修饰侧写”技术应用于特定生物学问题的研究来证明它们的有效性。错误调节的酪氨酸磷酸化是许多类型癌症的一个特征，有几种成功的抗癌疗法被设计来抑制与特定癌症相关的激活的酪氨酸激酶。因此，更多与癌症相关的酪氨酸激酶活性例子的知识最终可以转化为新的临床疗法。这个项目最初将允许候选人在赞助商David Eisenberg博士以及合作者Joseph Loo博士和Charles Sawyers博士的指导下进行学术发展，并最终将导致一个独立的研究项目。本项目将开发的收集蛋白质修饰数据的方法将补充现有的测量基因表达数据的方法，有助于实现基因组学和蛋白质组学的基本目标之一--了解细胞的电路。

DESCRIPTION: (provided by applicant): The cellular signaling circuitry has many types of regulation, from transcriptional control to regulation of protein degradation. These processes are often modulated by enzymatic modification of the proteins involved, and thus the modification of proteins is an integral part of most signal transduction pathways. In this project we plan to develop methods for detecting modifications within the proteome. Our initial focus will be on detecting tyrosine phosphorylation of proteins in cancer. We plan to develop methods for enriching cellular extracts for modified proteins directly coupled to methods for mass spectrometry-based detection and identification of these proteins. We also plan to demonstrate the usefulness of these "modification profiling" techniques by applying them to the study of specific biological questions. Misregulated tyrosine phosphorylation is a characteristic of many types of cancer, and several successful anticancer therapies are designed to inhibit activated tyrosine kinases associated with particular cancers. Thus, knowledge of additional examples of tyrosine kinase activities linked to cancer can ultimately be translated into new clinical therapies. This project will initially allow for the scholarly development of the candidate under the mentorship of the sponsor Dr. David Eisenberg, and the collaborators Dr. Joseph Loo and Dr. Charles Sawyers, and will ultimately result in an independent research project. The methods for collecting protein modification data to be developed in this project will complement existing methods for measuring gene expression data, assisting in one of the fundamental goals of genomics and proteomics - to understand the circuitry of the cell.