S-nitrosylation of A -adrenergic receptors inthe lung

描述（由申请人提供）： 该应用程序旨在使主要研究者能够开发 作为肺血管领域的独立医生-科学家， 生物学 在两位资深研究者的共同指导下， 乔纳森·斯塔姆勒和罗伯特·莱夫科维茨医生私家侦探将获得新的专业知识， 肺血管中α-肾上腺素能受体功能的研究， 大大加强了她在NO生物化学和蛋白质方面专业知识水平 通过亚硝基化修饰。 初步数据显示， 由α-肾上腺素能激动剂引起的血管收缩被内皮- 衍生舒张因子一氧化氮（NO）。 NO（或其衍生分子） 已知通过半胱氨酸的化学修饰来调节蛋白质功能 残基，即亚硝基化。 具体的目标测试的假设，没有 通过化学修饰α 1 β-肾上腺素能调节肺血管张力 肺血管平滑肌中的受体。 （1）确定哪些 α 1-肾上腺素能受体信号传导的组分被NO修饰， 拮抗血管收缩。 目的2）确定NO S-亚硝基化是否 （a1-肾上腺素能受体或Aim 1中检测到的其他靶点，并确定 半胱氨酸参与。 目的3）研究的作用和机制 组成型和诱导型一氧化氮合酶亚型在调节 体外和体内a1肾上腺素能功能。 实验将在 用α 1 β-肾上腺素能受体转染的HEK 293（目的1和2），培养大鼠 肺血管细胞和肺动脉环（Aim 3）。 该提案还将 确定NO的作用是否特异于α 1-肾上腺素能 受体或代表NO调节G蛋白偶联的一般机制 受体信号 了解调节肺功能的因素 血管功能将导致肺新疗法的合理开发 影响肺血管的疾病。

DESCRIPTION (provided by applicant): This application is designed to enable the principal investigator to develop as an independent physician-scientist in the field of pulmonary vascular biology. Under the co-mentorship of two established investigators, Dr. Jonathan Stamler and Dr. Robert Lefkowitz, the P.I. will gain new expertise in the study of alpha-adrenergic receptor function in the pulmonary vasculature and substantially strengthen her level of expertise in NO biochemistry and protein modification by Snitrosylation. Preliminary data show that pulmonary vasoconstriction by a-adrenergic agonists is attenuated by the endothelial- derived relaxing factor, nitric oxide (NO). NO (or molecules derived from it) is known to regulate protein function by chemical modifications of cysteine residues, i.e. Snitrosylation. The specific aims test the hypothesis that NO regulates pulmonary vascular tone by chemically modifying alpha1Beta-adrenergic receptors in pulmonary vascular smooth muscle. Aim 1) Determine which components of (alpha1-adrenergic receptor signaling are modified by NO to antagonize vasoconstriction. Aim 2) Determine whether NO S-nitrosylates the (alpha1-adrenergic receptor or other targets detected in Aim 1, and identify the cysteines involved. Aim 3) Investigate the role and mechanisms of constitutive and inducible nitric oxide synthase isoforms in the regulation of a1-adrenergic function in vitro and in vivo. Experiments will be performed in HEK293 transfected with alpha1beta-adrenergic receptor (Aims 1 and 2), cultured rat pulmonary vascular cells and PA rings (Aim 3). This proposal will also establish whether the effects of NO are specific for the (alpha1-adrenergic receptor or represent a general mechanism for NO to regulate G-protein coupled receptor signaling. Understanding the factors that regulate pulmonary vascular function will lead to rational development of new therapies for lung diseases that affect the pulmonary blood vessels.