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（NOS 3）和p38α在细胞内信号传导中起关键作用。 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，非常接近信号传导的末端， 在体外和内皮细胞的环境中，它级联地在NOS 3中起作用并与NOS 3结合。 细胞NOS 3是血管稳态的关键调节因子，并对各种各样的 信号（例如，缓激肽、VEGF、胰岛素）通过产生一氧化氮（NO），其本身是一种重要的 信号分子NOS 3的调节通过蛋白质相互作用发生， 翻译后修饰、细胞定位和支架。的成果 p38α与NOS 3的结合仍然是个谜，但结合在细胞内是受调节的。 环境对氧化的调节作用的新认识和 激活剂和底物募集对MAPK的作用使我们假设， 也许p38α-NOS 3结合的关键作用是允许NOS 3通过 支架化以控制基底接近和/或通过氧化直接抑制。我们提出 采用邻位连接法和生物层干涉法， 变体）和NOS 3（或NOS 3肽），以研究这种变化的动态性质。 同时使用体外和原位分析与内皮细胞的相互作用来测试我们的假设， 细胞这个R15区域项目将提供有关以下交叉点的重要信息： NOS 3和p38，同时涉及和暴露本科研究人员目前 细胞信号的问题，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.