Characterization of AQP2 interacting proteins and novel functions of AQP2

AQP2 相互作用蛋白的表征和 AQP2 的新功能

• 批准号: 7138395
• 负责人: HUA A LU
• 金额: $ 13.43万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-05 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7138395
• 关键词: cell adhesion; cell cell interaction; cell migration; chimeric proteins; endocytosis; exocytosis; heat shock proteins; laboratory mouse; phosphorylation; protein binding; protein protein interaction; protein structure function; protein transport; renal tubule; small interfering RNA; tissue /cell culture; transfection; water channel; zebrafish

DESCRIPTION (provided by applicant): This applicant has proposed a program of research to prepare her for a career in academic nephrology and basic science research in the field of renal physiology/pathophysiology, specifically, AQP2 trafficking. This applicant will propose to characterize AQP2 interacting proteins and investigate the physiological significance of these interactions, to better understand the molecular mechanism underlying AQP2 trafficking, and explore the possible novel role of AQP2 in cellular biological and pathophysiological processes. The research will be conducted in the laboratory of Dr. Dennis Brown at the Program in Membrane Biology (PMB) and Division of Nephrology, Massachusetts General Hospital. Vasopressin (VP) is the major antidiuretic hormone involved in the regulation of water reabsorption by mammalian kidney. It 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 fluid retention seen in congestive heart failure, cirrhosis, as well as concentrating defect seen in diabetes insipidus. AQP2 is regulated through complex trafficking pathways which have not been well characterized. Our hypothesis is that regulated trafficking of AQP2 requires direct and indirect protein-protein interactions during intracellular translocation, exocytosis as well as endocytosis. Specifically, we will 1) extend our current study on the interaction of AQP2 and heat shock protein 70 (hsc70) by tracking further down to subcellular compartments/step(s) in AQP2 trafficking pathways, and continue to characterize other AQP2 binding candidates identified from our yeast-two hybrid screen; 2) study the molecular mechanism underlying the role of phosphorylation and dephosphorylation on AQP2 trafficking; 3) explore novel potential roles of AQP2 in cell adhesion, migration, and renal tubular formation/malformation. We will investigate the distribution and trafficking of AQP2 during the general biological processes as well as the effect of alteration of AQP2 trafficking on these biological processes. We will using stable or primary cell culture, tissue slice culture, 3D culture system, a zebrafish model system, and AQP2 mutant mouse model. Multidisciplinary approaches and powerful technologies will be used to carry out these studies that will shed light on the mechanism of AQP2 trafficking and function.

描述（由申请人提供）： 该申请人提出了一项研究计划，为她在肾脏生理学/病理生理学领域（特别是 AQP2 贩运）的学术肾病学和基础科学研究的职业生涯做好准备。该申请人将提出表征AQP2相互作用蛋白并研究这些相互作用的生理意义，以更好地理解AQP2运输的分子机制，并探索AQP2在细胞生物学和病理生理学过程中可能的新作用。该研究将在马萨诸塞州总医院膜生物学项目 (PMB) 和肾内科 Dennis Brown 博士的实验室中进行。加压素 (VP) 是参与哺乳动物肾脏水重吸收调节的主要抗利尿激素。它通过将 AQP2 水通道从细胞质囊泡募集到集合管主细胞的质膜来发挥作用。 VP-AQP2 信号通路受损会导致充血性心力衰竭、肝硬化中出现的液体潴留，以及尿崩症中出现的集中缺陷。 AQP2 通过复杂的运输途径进行调节，但这些途径尚未得到充分表征。我们的假设是，AQP2 的调节运输需要细胞内易位、胞吐作用和内吞作用期间直接和间接的蛋白质-蛋白质相互作用。具体来说，我们将 1) 通过进一步追踪 AQP2 运输途径中的亚细胞区室/步骤，扩展我们目前关于 AQP2 和热休克蛋白 70 (hsc70) 相互作用的研究，并继续表征从我们的酵母-两种杂交筛选； 2）研究磷酸化和去磷酸化对AQP2运输作用的分子机制； 3）探索AQP2在细胞粘附、迁移和肾小管形成/畸形中的新的潜在作用。我们将研究 AQP2 在一般生物过程中的分布和运输，以及 AQP2 运输的改变对这些生物过程的影响。我们将使用稳定或原代细胞培养、组织切片培养、3D 培养系统、斑马鱼模型系统和 AQP2 突变小鼠模型。将使用多学科方法和强大的技术来开展这些研究，以阐明 AQP2 运输和功能的机制。