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：
6390243
负责人：
RICHARD C VENEMA
金额：
$ 22.17万
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/6390243
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.

内皮型和诱导型一氧化氮合酶（eNOS 和 iNOS）在正常血管生理学和血管病理学中都具有关键的调节作用。该提案中提出的初步研究表明，eNOS 直接与缓激肽 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诱导。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 也在平滑肌中相互作用。在此应用中，我们建议检验 eNOS 在内皮细胞中通过与 B2 受体的抑制性相互作用进行调节的假设。我们还将确定酪氨酸磷酸化在相互作用调节中的作用，并定义负责相互作用的两种蛋白质中的结构域。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释放 NO，并将确定这两种蛋白质中的相互作用域。最后，我们将测试 iNOS-CAT2 相互作用促进血管平滑肌细胞释放 NO 的假设，并将定义参与相互作用的两种蛋白质的结构域。