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

NOS3 和 p38 MAP 激酶 - 它们之间的相互作用是 p38 调节机制吗？

• 批准号: 10653595
• 负责人: Carol Ann Chrestensen
• 金额: $ 40.57万
• 依托单位: KENNESAW STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10653595
• 关键词: Affect; Affinity; Atherosclerosis; Binding; Binding Sites; Biological; Biological Assay; Blood Vessels; Bradykinin; Cells; Cysteine; Diabetes Mellitus; Dimerization; Disease; Endothelial Cells; Environment; Enzymes; Escherichia coli; Event; Family; Family member; Homeostasis; In Situ; In Vitro; Insulin; Interferometry; Ligation; Location; MAPK8 gene; Malignant Neoplasms; Mitogen-Activated Protein Kinases; Modification; Molecular; N-terminal; NOS1 gene; NOS2A gene; NOS3 gene; Nature; Nitric Oxide; Nitric Oxide Synthase; Outcome; Output; PKA inhibitor; Pathway interactions; Peptides; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Physiological Processes; Play; Post-Translational Protein Processing; Process; Production; Protein Kinase Interaction; Proteins; Regulation; Research; Research Personnel; Rest; Role; Running; SRC gene; STEM field; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Stimulus; Surface; Synaptic Transmission; Testing; Variant; Vascular Endothelial Growth Factors; Vasodilation; career; dimer; enzyme activity; experimental study; immune function; in vitro activity; inhibitor; member; nanomolar; oxidation; p38 Mitogen Activated Protein Kinase; protein protein interaction; recruit; response; scaffold; success; time use; undergraduate student

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.

项目摘要 一氧化氮合酶3（NOS3）和p38α在细胞内信号传导中起关键作用。 这些酶的活性的改变会影响诸如动脉粥样硬化的疾病， 糖尿病和癌症。 p38无处不在，非常接近信号的末端 级联在体外和内皮环境中发挥作用并与NOS3结合 细胞。 NOS3是血管稳态的关键调节剂，对各种各样的反应 通过产生一氧化氮（NO）本身的信号（例如Bradykinin，VEGF，胰岛素）本身就是重要的 信号分子。 NOS3的调节是通过蛋白质相互作用发生的， 翻译后修饰，细胞定位和脚手架。结果 p38α与NOS3的结合仍然是神秘的，但在细胞中调节结合 环境。对氧化和关键的调节作用的新兴理解 激活因子和底物在MAPK上扮演的角色使我们假设 p38α-NOS3结合的关键作用可能是允许NOS3通过 脚手架以控制底物进入和/或通过氧化直接抑制。我们建议 使用接近连接测定法和对p38α（野生类型和 变体）和NOS3（或NOS3 Pepperides），以研究此的动态性质 在使用体外和原位分析测试我们的假设的同时与内皮进行现场分析 细胞。这个R15区域项目将提供有关相交的重要信息 NOS3和P38涉及并将本科研究人员暴露于当前 细胞信号传导中的问题，为它们在茎场中的职业做准备。