Profiling of Protein Modifications by Mass Spectrometry

产品说明：（申请人提供）：细胞信号通路有许多类型的调节，从转录控制到蛋白质降解的调节。这些过程通常通过所涉及的蛋白质的酶促修饰来调节，因此蛋白质的修饰是大多数信号转导途径的组成部分。在这个项目中，我们计划开发用于检测蛋白质组中的修饰的方法。我们最初的重点将是检测癌症中蛋白质的酪氨酸磷酸化。我们计划开发用于富集修饰蛋白质的细胞提取物的方法，该方法直接与用于基于质谱的检测和鉴定这些蛋白质的方法相结合。我们还计划通过将这些“修饰分析”技术应用于特定生物学问题的研究来证明它们的有用性。酪氨酸磷酸化失调是许多类型癌症的特征，并且几种成功的抗癌疗法被设计为抑制与特定癌症相关的活化酪氨酸激酶。因此，与癌症相关的酪氨酸激酶活性的其他实例的知识最终可以转化为新的临床治疗。该项目最初将允许候选人在赞助商大卫艾森伯格博士以及合作者约瑟夫卢博士和查尔斯索耶博士的指导下进行学术发展，并最终导致一个独立的研究项目。该项目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.