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