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

项目概要 一氧化氮合酶 3 (NOS3) 和 p38α 在细胞内信号传导中发挥关键作用。 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 在体外和内皮细胞环境中，它在 NOS3 中发挥作用并与 NOS3 结合 细胞。 NOS3 是血管稳态的关键调节因子，可对多种因素做出反应 通过产生一氧化氮 (NO) 来释放信号（例如缓激肽、VEGF、胰岛素），一氧化氮本身是一种重要的 信号分子。 NOS3 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。结果 p38α 与 NOS3 的结合仍然是个谜，但结合在细胞中受到调节 环境。对氧化调节作用及其关键的新认识 激活剂和底物招募对 MAPK 的作用使我们推测 也许 p38α-NOS3 结合的一个关键作用是允许 NOS3 通过以下方式调节 p38α： 控制底物进入和/或通过氧化直接抑制的支架。我们建议 使用 p38α（野生型和 变体）和 NOS3（或 NOS3 肽），以研究其动态性质 相互作用，同时使用内皮细胞的体外和原位分析来测试我们的假设 细胞。这个 R15 AREA 项目将提供有关交叉点的重要信息 NOS3 和 p38 同时让本科生研究人员接触和接触当前的 细胞信号传导问题，为他们在 STEM 领域的职业生涯做好准备。

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.