NOS3 and p38 MAP kinase - is the interaction between them a mechanism of p38 regulation?

项目摘要 一氧化氮合酶3(NOS3)和p38α在细胞内信号转导中起关键作用。 这些酶活性的改变会影响动脉粥样硬化等疾病， 糖尿病和癌症。P38普遍表达，非常接近信号的末尾。 在体外和内皮环境中，它级联起作用并与NOS3结合 细胞。NOS3是血管动态平衡的关键调节器，对多种不同的 信号(例如，缓激肽、血管内皮生长因子、胰岛素)通过产生一氧化氮(NO)本身是一种重要的 信号分子。NOS3的调节是通过蛋白质的相互作用进行的， 翻译后修饰、细胞定位和支架。其结果是 P38α与nos3的结合仍然是个谜，但结合在细胞内是被调节的。 环境。对氧化的调节作用和关键的新认识 激活剂和底物招募对MAPK的作用使我们假设 也许p38α-nos3结合的一个关键作用是允许nos3通过 脚手架，以控制底物的访问和/或通过氧化直接抑制。我们建议 用p38α(野生型和 变异体)和NOS3(或NOS3肽)，以研究其动态性质 使用体外和原位内皮分析来验证我们的假设时的相互作用 细胞。该R15区域项目将提供有关 NOS3和p38，同时让本科生研究人员接触到当前的 细胞信号方面的问题，为他们在STEM领域的职业生涯做准备。

Project Summary Nitric Oxide Synthase 3 (NOS3) and p38α play key roles in intracellular signaling. Alterations to the activity of these enzymes impact diseases like atherosclerosis, diabetes, and cancer. p38 is ubiquitously expressed, very near the end of the signaling cascades it functions in and bind to NOS3 in vitro and in the environment of endothelial cells. NOS3 is a key regulator of vascular homeostasis and responds to a wide variety of signals (e.g., bradykinin, VEGF, insulin) by producing nitric oxide (NO), itself an important signaling molecule. Regulation of NOS3 occurs through protein interactions, posttranslational modifications, cellular localization, and scaffolding. The outcomes of p38α binding to NOS3 are still mysterious, yet binding is modulated in the cellular environment. Emerging understanding of the regulatory roles of oxidation and the key role that activator and substrate recruitment has on MAPKs led us to hypothesize that perhaps a key role of p38α-NOS3 binding is to allow NOS3 to regulate p38α through scaffolding to control substrate access and/or direct inhibition by oxidation. We propose to use proximity ligation assay and biolayer interferometry with p38α (wild type and variants) and NOS3 (or NOS3 peptides), to investigate the dynamic nature of this interaction while testing our hypotheses using in vitro and in situ analysis with endothelial cells. This R15 AREA project will provide important information about the intersection of NOS3 and p38 while involving and exposing undergraduate researchers to current questions in cellular signaling, preparing them for careers in STEM fields.