Characterization of Aquaporin 2 Interacting Proteins

18. 申请者姓名（最后一名，机构贝林医科大学马萨诸塞大学第一名，中间首字母）导师陆兆峰博士，M.D. Dr. John Burand博士，Dr. Dr. John Burand博士，博士机构/公司主管/雇主University . Massachusetts University . J. Burand BIDMC, HMS Dr. V. Sukhatme NYUMC Dr. R. Raitch NYUMC Dr. M. Charap MGH，本申请中提出的研究将在我在麻省总医院的三年博士后研究期间进行（2002年7月- 2005年6月）。研究结束后，我将在同一机构完成一年的临床肾脏病学研究。我在病毒学和分子生物学方面受过广泛的训练。通过加入Dr. Dennis Brown的实验室，我非常有信心我将接触到一个优秀的科学环境，在那里我将学习到令人兴奋的肾脏生理学和病理生理学领域。特别是，我将在膜生物学/蛋白质运输方面获得额外的培训。我相信这三年的奖学金培训将为我提供扎实的科学知识和丰富的经验，为我作为独立研究者的职业生涯奠定坚实的基础。我也把这个研究奖学金视为我教育的一个组成部分，因为我的目标是在学术肾脏病学中追求职业生涯，在那里我可以结合病人护理，研究和教学。我_“佐薇]_F, 1;e]我19岁。姓名及学位：丹尼斯·布朗，博士。职务/职级肾科21副主任。研究兴趣/领域血管加压素（VP）是一种主要的抗利尿激素，参与调节哺乳动物肾脏的水重吸收。大量研究表明，VP通过将AQP2水通道从细胞质囊泡募集到集管主细胞的质膜来发挥作用。VP-AQP2信号通路的损伤导致尿崩症和一些心力衰竭患者的液体潴留。AQP2通过复杂的贩运途径受到调节。我们的假设是，AQP2的调节运输需要在细胞内易位、胞吐和内吞过程中直接和间接的蛋白质-蛋白质相互作用。拟议的研究分为三个具体目标，分别是鉴定和表征新的AQP2相互作用蛋白，以及研究这些新的蛋白相互作用对体外和体内AQP2运输的功能意义。该研究将利用多学科方法（分子生物学、细胞生物学、生物化学和免疫细胞化学）和强大的技术（酵母双杂交筛选、结构功能分析、基因转移）来鉴定和表征新的蛋白质-蛋白质相互作用及其生物学功能。这些研究将为尚不清楚的AQP2转运的分子机制提供重要信息，并可能最终为尿浓度缺陷患者提供更好的治疗方法。小灵通416 - 1(启06/02)表单页面2 Kirschstein-NRSA个人奖学金申请NAMEOFAPPLICANT(最后,首先,中间初始)表的内容 ======================================== 节结束 ===========================================

18. GOALS FOR KIRSCHSTEIN-NRSA FELLOWSHIP TRAINING AND CAREER Lu, Hua, A. J. I NAME OF APPLICANT (Last, INSTITUTION Belling Medical University University of Massachusetts University of Massachusetts first, middle initial) MENTOR Dr. Zhao-Feng Lu, M.D. Dr. John Burand, Ph.D. Dr. John Burand, Ph.D INSTITUTION/COMPANY SUPERVISOR/EMPLOYER Univ. Massachus Dr. J. Burand Univ. Massachus Dr. J. Burand BIDMC, HMS Dr. V. Sukhatme NYUMC Dr. R. Raitch NYUMC Dr. M. Charap MGH, HMS Dr. D. Brown The research proposed in this application will be conducted during my three years of postdoctoral fellowship (July 2002-Jun 2005) at the Massachusetts General Hospital. Immediately after the research years, I am going to complete one year of clinical nephrology fellowship at the same institution. I have extensive training in virology and molecular biology. By joining Dr. Dennis Brown's laboratory, I am very confident that I will be exposed to an excellent scientific environment, where I will learn about the exciting field of renal physiology and pathophysiology. In particular, I will gain additional training in membrane biology/protein trafficking. I believe that this three years' fellowship training will provide me with solid scientific knowledge and extensive experiences to build a strong foundation for a career as an independent investigator. I also see this research fellowship as an integral part of my education, as my goal is to pursue a career in academic nephrology, where I could combine patient care, research and teaching. I _"-zo]_F,1;e]i 19. NAME AND DEGREE(S) Dennis Brown, PhD 20. POSITION/RANK Associate Chief, Renal Unit 21. RESEARCH INTERESTS/AREAS 22. DESCRIPTION Vasopressin (VP) is the major antidiuretic hormone involved in the regulation of water reabsorption by mammalian kidney. Intensive studies have established that VP functions by recruiting the AQP2 water channel from cytoplasmic vesicles to the plasma membrane of collecting duct principal cells. The impairment of VP-AQP2 signaling pathways results in diabetes insipidus as well as fluid retention seen in some heart failure patients. AQP2 is regulated through complex trafficking pathways. Our hypothesis is that regulated trafficking of AQP2 requires direct and indirect protein-protein interactions during intracellular translocation, exocytosis as well as endocytosis. The proposed studies are divided into three specific aims addressing identification and characterization of novel AQP2 interacting proteins as well as investigation of the functional significance of these novel protein interactions on AQP2 trafficking in vitro and in vivo. The proposed study will utilize multidisciplinary approaches (molecular biology, cell biology, biochemistry and immunocytochemistry) and powerful technology (yeast two hybrid screen, structural functional analysis, gene transfer) to identify and characterize novel protein- protein interactions and their biologic functions. These studies will provide important information of the poorly understood molecular mechanisms underlying AQP2 trafficking, and may ultimately lead to better therapies for patients with defective urinary concentration. PHS 416-1 (Rev. 06/02) Form Page 2 Kirschstein-NRSA Individual Fellowship Application NAMEOFAPPLICANT(Last,first, middle initial) Table of Contents ========================================Section End===========================================