D. J. Burke,
H. E. Boudreau,
K. J. Lekstrom,
D. R. Ambruso,
S. H. Jackson, and
T. L. Leto
Free Radical Biology and Medicine,
Nox1 is an abundant source of reactive oxygen species (ROS) in colon epithelium recently shown to function in wound healing and epithelial homeostasis. We identified Peroxiredoxin 6 (Prdx6) as a novel binding partner of Nox activator 1 (Noxa1) in yeast two-hybrid screening experiments using the Noxa1 SH3 domain as bait. Prdx6 is a unique member of the Prdx antioxidant enzyme family exhibiting both glutathione peroxidase and phospholipase A2 activities. We confirmed this interaction in cells overexpressing both proteins, showing Prdx6 binds to and stabilizes wild type Noxa1, but not the SH3 domain mutant form, Noxa1 W436R. We demonstrated in several cell models that Prdx6 knockdown suppresses Nox1 activity, whereas enhanced Prdx6 expression supports higher Nox1-derived superoxide production. Both peroxidase- and lipase-deficient mutant forms of Prdx6 (Prdx6 C47S and S32A, respectively) failed to bind to or stabilize Nox1 components or support Nox1-mediated superoxide generation. Furthermore, the transition-state substrate analogue inhibitor of Prdx6 phospholipase A2 activity (MJ-33) was shown to suppress Nox1 activity, suggesting Nox1 activity is regulated by the phospholipase activity of Prdx6. Finally, wild type Prdx6, but not lipase or peroxidase mutant forms, supports Nox1-mediated cell migration in the HCT-116 colon epithelial cell model of wound closure. These findings highlight a novel pathway in which this antioxidant enzyme positively regulates an oxidant-generating system to support cell migration and wound healing.
author = "J. Kwon and A. Wang and D. J. Burke and H. E. Boudreau and K. J. Lekstrom and A. Korzeniowska and R. Sugamata and Y.-S. Kim and L. Yi and I. Ersoy and S. Jaeger and K. Palaniappan and D. R. Ambruso and S. H. Jackson and T. L. Leto",
title = "Peroxiredoxin 6 (Prdx6) supports NADPH oxidase1 (Nox1)-based superoxide generation and cell migration",
year = 2016,
journal = "Free Radical Biology and Medicine",
month = "Apr",
keywords = "cell migration, biomedical",
doi = "10.1016/j.freeradbiomed.2016.04.009"
J. Kwon, A. Wang, D. J. Burke, H. E. Boudreau, K. J. Lekstrom, A. Korzeniowska, R. Sugamata, Y.-S. Kim, L. Yi, I. Ersoy, S. Jaeger, K. Palaniappan, D. R. Ambruso, S. H. Jackson, and T. L. Leto. Peroxiredoxin 6 (Prdx6) supports NADPH oxidase1 (Nox1)-based superoxide generation and cell migration. Free Radical Biology and Medicine, April 2016.