Structure and function of integrin TM and CYTO domains

描述(由申请人提供)：应聘者的职业目标是成为一名独立的研究员，重点研究整合素介导的信号通路的分子机制，并采用跨学科的方法。许多整合素与细胞发育和迁移密切相关，因此被认为是治疗癌症的潜在药物靶点。在短期内，候选人寻求了解整合素αIIbbeta3的跨膜和细胞质(TM-Cyto)结构域的作用，使用结构和细胞生物学方法相结合。这项研究的结果将有助于更好地了解癌症生物学和抗癌药物的开发。为此，他启动了与AlphaIIb和Beta3TM-Cyto结构域相对应的膜结合蛋白的研究，并假设这些蛋白的同源寡聚化在整合素激活和聚集的背景下的意义。本项目的具体目标是：(1)用核磁共振波谱确定β3亚基TM-Cyto结构域的溶液结构。(2)鉴定生物膜中？IIB和？3TM-Cyto结构域的同源齐聚，并鉴定破坏或增强自结合的点突变。基于细菌的TOXCAT系统将用于这一目的。(3)研究同源寡聚对GM1500 B淋巴细胞和/或中国仓鼠卵巢细胞αIIbbeta3功能的影响。对于候选人来说，未来一两年将是关键的一年，他将提高自己的研究技能，并在整合素领域做出重大贡献，从而使其能够平稳过渡到独立。他的赞助人威廉·德格拉多博士有着出色的指导记录。在赞助商的实验室和生物化学与生物物理系，有一种支持和激励的氛围，鼓励科学交流和合作，这有利于候选人作为一名科学家的发展。

DESCRIPTION (provided by applicant): The candidate's career goal is to become an independent investigator focused on the molecular mechanisms of the integrin-mediated signaling pathway with an interdisciplinary approach. Many integrins are critically involved in cell development and migration, and accordingly identified as potential drug targets for cancer treatment. In the short term, the candidate seeks to understand the roles of the transmembrane and cytoplasmic (TM-CYTO) domains of integrin alphaIIbbeta3, using a combination of structural and cell biology methods. The results acquired from this study will contribute to a better understanding of cancer biology and the development of anti-cancer agents. To this end, he has initiated studies on the membrane-bound proteins corresponding to alphaIIb and beta3TM-CYTO domains, and hypothesized the significance of homo-oligomerization of these proteins in the context of integrin activation and clustering. The specific aims of this project are: (1) To determine the solution structure of the TM-CYTO domains of ?3 subunit by NMR spectroscopy. (2) To characterize homo-oligomerization of the ?IIb and ?3TM-CYTO domains in biological membranes and identify point mutations that disrupt or enhance self-association. The bacteria-based TOXCAT system will be used for this purpose. (3) To study the effects of homo-oligomerization on alphaIIbbeta3 function in GM1500 B-lymphocytes and/or Chinese hamster ovary cells. The next year or two will be critical for the candidate to polish his research skills, and make a significant contribution to the integrin field, thus allowing for a smooth transition to independence. His sponsor, Dr. William DeGrado, has an excellent mentoring track record. There is a supportive and stimulating atmosphere in the sponsor's lab and the Department of Biochemistry and Biophysics that encourages scientific exchange and collaboration, which is beneficial to the candidate's development as a scientist.