RECEPTOR AND TRANSPORTER REGULATION OF NO SYNTHASES

无合酶的受体和转运蛋白调节

基本信息

批准号：
6184598
负责人：
RICHARD C VENEMA
金额：
$ 21.53万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-07-01 至 2003-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6184598
关键词：
aminoacid transport binding sites enzyme induction /repression enzyme mechanism enzyme structure luminescence membrane transport proteins neuropeptide receptor nitric oxide nitric oxide synthase phosphorylation protein protein interaction receptor binding tissue /cell culture transfection vascular endothelium vascular smooth muscle yeast two hybrid system

项目摘要

The endothelial and inducible nitric oxide synthases (eNOS and iNOS) have key regulatory roles in both normal vascular physiology and in vascular pathology. Preliminary studies presented in this proposal indicate that eNOS interacts directly with the bradykinin B2 receptor and with the CAT1 cationic amino acid transporter in vascular endothelial cells. In addition, it has been shown that iNOS is coinduced with the CAT2 cationic amino acid transporter in vascular smooth muscle cells. Because iNOS and CAT2 are structurally and functionally very similar to eNOS and CAT1, we postulate that iNOS and CAT2 also interact in smooth muscle. In this application, we propose to test the hypothesis that eNOS is regulated in endothelial cells by inhibitory interactions with the B2 receptor. We will also determine the role of tyrosine phosphorylation in regulation of the interaction and will define the domains in the two proteins that are responsible for the interaction. We further propose to test the hypothesis that the eNOS-CAT1 interaction facilitates NO release from endothelial cells and will determine what the interacting domains are in the two proteins. Finally, we will test the hypothesis that the iNOS-CAT2 interaction facilitates NO release from vascular smooth muscle cells and will define the domains in the two proteins that are involved in the interaction.

内皮和诱导的一氧化氮合酶（ENOS和INOS）在正常血管生理和血管病理学中具有关键的调节作用。该提案中介绍的初步研究表明，eNOS直接与Bradykinin B2受体以及血管内皮细胞中的CAT1阳离子氨基酸转运蛋白相互作用。此外，已经表明，iNOS与血管平滑肌细胞中的Cat2阳离子氨基酸转运蛋白共同引起。由于iNOS和CAT2在结构和功能上与eNOS和CAT1非常相似，因此我们假设Inos和Cat2在平滑肌中也相互作用。在此应用中，我们建议检验以下假设：eNOS通过与B2受体抑制性相互作用在内皮细胞中受到调节。我们还将确定酪氨酸磷酸化在相互作用调节中的作用，并将定义两种负责相互作用的域中的结构域。我们进一步建议测试以下假设：eNOS-CAT1相互作用促进内皮细胞没有释放，并将确定两种蛋白质中的相互作用域是什么。最后，我们将检验以下假设：iNOS-CAT2相互作用促进了血管平滑肌细胞没有释放，并将定义相互作用中涉及的两种蛋白质中的结构域。