Functional Interaction of ANP and NO signaling

ANP 和 NO 信号传导的功能相互作用

• 批准号: 7632226
• 负责人: KUMAR U KOTLO
• 金额: $ 13.82万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7632226
• 关键词: Acute Kidney Failure; Affinity Chromatography; Area; Atrial Natriuretic Factor; Atrial Natriuretic Factor Receptors; Basic Science; Blood Pressure; Canis familiaris; Cell Culture Techniques; Cell physiology; Cells; Chicago; Chronic; Chronic Kidney Failure; Clinical; Complex; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Cytoplasm; Data; Disease; Environment; Enzymes; Epithelial Cells; Family; Fluorescent Antibody Technique; Goals; Guanosine Triphosphate; Guanylate Cyclase; Heme; Human; Hypertension; Illinois; Immunohistochemistry; Immunoprecipitation; In Vitro; Kidney; Kidney Diseases; Knowledge; Laboratory Research; Measures; Mediating; Medicine; Membrane; Methods; Molecular; Muscle relaxation phase; Nitric Oxide; Physiological; Platelet aggregation; Protein Isoforms; Proteins; RNA Interference; Receptor Signaling; Recombinants; Regulation; Renal glomerular disease; Renal tubule structure; Research; Research Personnel; Role; Signal Pathway; Signal Transduction; Smooth Muscle; Sodium; Technology; Training; Tubular formation; Universities; base; career; design; experience; extracellular; in vivo; kidney epithelial cell; novel; phosphodiesterase V; phosphoric diester hydrolase; protein complex; protein protein interaction; skills; therapeutic target; uptake

DESCRIPTION (provided by applicant): The current proposal is aimed at providing the PI with extensive training in receptor signaling with reference to its clinical implications along with formal courses at the graduate level to enhance his research and laboratory skills. The experienced sponsor and co-sponsor, along with a highly interactive basic-research environment at department of Medicine at the University of Illinois at Chicago, offer a great training opportunity for the PI to achieve his career goals in becoming an independent research investigator in the area of the signaling mechanisms. The proposed studies are aimed at examining the molecular and functional interaction of atrial natriuretic peptide (ANP) and nitric oxide (NO) signaling. Both ANP and NO increase intracellular cyclic GMP (cGMP) levels thus leading to reduction in arterial blood pressure and inhibition of renal tubular sodium transport. Studies in specific Aim 1 will determine the interacting domains of NOGCb1 and GCA utilizing immunoprecipitations and immunofluorescence techniques. We will also identify proteins interacting with GCA and NOGCb1 , possibly novel ones by using novel Tandem Affinity Purification (TAP) method. In specific Aim 2, we will focus on evaluating the functional impact of the association of NOGCb1 with GCA on ANP-mediated stimulation of cGMP, cGMP-dependent protein kinase (cGK), and cGMP phosphodiesterase (PDE) using RNAi technology. In specific Aim 3 we will determine the effect of reducing NOGCb1 on ANP signaling in renal tubular sodium handling by measuring 22Na uptake in renal epithelial cells transfected with RNAi against NOGCb1 and treated with ANP. The functional interaction between ANP and NO signaling may provide new understanding to diseases associated with reduced NO signaling, such as renal glomerular disease, chronic and acute renal failure, and hypertension.

描述（由申请人提供）： 当前的建议旨在为PI提供有关受体信号传导的广泛培训，并参考其临床意义以及研究生层面的正式课程，以增强其研究和实验室技能。这位经验丰富的赞助商和共同赞助商，以及伊利诺伊大学芝加哥大学医学系的高度互动的基础研究环境，为PI提供了很大的培训机会，以实现他的职业目标，以成为信号机制领域的独立研究研究员。拟议的研究旨在检查房屋亚催化肽（ANP）和一氧化氮（NO）信号的分子和功能相互作用。 ANP和无增加的细胞内循环GMP（CGMP）水平，从而导致动脉血压降低和抑制肾小管钠转运。特定目标1中的研究将确定利用免疫沉淀和免疫荧光技术的NOGCB1和GCA的相互作用结构域。我们还将通过使用新颖的串联亲和力纯化（TAP）方法来鉴定与GCA和NOGCB1相互作用的蛋白质。在特定目标2中，我们将重点介绍NOGCB1与GCA的关联对ANP介导的CGMP刺激CGMP，CGMP依赖性蛋白激酶（CGK）和CGMP磷酸二酯酶（PDE）的功能影响。在特定的目标3中，我们将通过测量用RNAi针对NOGCB1转染的肾上皮细胞中的22NA摄取并用ANP处理，从而确定减少NOGCB1对肾小管钠处理中ANP信号传导的影响。 ANP和NO信号之间的功能相互作用可能会对与降低没有信号传导相关的疾病（例如肾肾小球疾病，慢性和急性肾功能衰竭以及高血压）提供新的理解。