Journal article
Phosphorylation of Nox1 regulates association with NoxA1 activation domain
Circulation research, Vol.115(11), pp.911-918
11/07/2014
DOI: 10.1161/CIRCRESAHA.115.304267
PMCID: PMC5025877
PMID: 25228390
Abstract
Activation of Nox1 initiates redox-dependent signaling events crucial in the pathogenesis of vascular disease. Selective targeting of Nox1 is an attractive potential therapy, but requires a better understanding of the molecular modifications controlling its activation.
To determine whether posttranslational modifications of Nox1 regulate its activity in vascular cells.
We first found evidence that Nox1 is phosphorylated in multiple models of vascular disease. Next, studies using mass spectroscopy and a pharmacological inhibitor demonstrated that protein kinase C-beta1 mediates phosphorylation of Nox1 in response to tumor necrosis factor-α. siRNA-mediated silencing of protein kinase C-beta1 abolished tumor necrosis factor-α-mediated reactive oxygen species production and vascular smooth muscle cell migration. Site-directed mutagenesis and isothermal titration calorimetry indicated that protein kinase C-beta1 phosphorylates Nox1 at threonine 429. Moreover, Nox1 threonine 429 phosphorylation facilitated the association of Nox1 with the NoxA1 activation domain and was necessary for NADPH oxidase complex assembly, reactive oxygen species production, and vascular smooth muscle cell migration.
We conclude that protein kinase C-beta1 phosphorylation of threonine 429 regulates activation of Nox1 NADPH oxidase.
Details
- Title: Subtitle
- Phosphorylation of Nox1 regulates association with NoxA1 activation domain
- Creators
- Jennifer Streeter - From the Departments of Internal Medicine (B.M.S., S.J., B.S., W.H.T., F.J.M.), Microbiology (J.C.D.H.), Anatomy and Cell Biology (J.S.), Biochemistry (L.G.), and Protein Crystallography Facility (L.G.), University of Iowa, Iowa City; and Veterans Affair Medical Center, Iowa City, IA (F.J.M.)Brandon M Schickling - From the Departments of Internal Medicine (B.M.S., S.J., B.S., W.H.T., F.J.M.), Microbiology (J.C.D.H.), Anatomy and Cell Biology (J.S.), Biochemistry (L.G.), and Protein Crystallography Facility (L.G.), University of Iowa, Iowa City; and Veterans Affair Medical Center, Iowa City, IA (F.J.M.)Shuxia Jiang - From the Departments of Internal Medicine (B.M.S., S.J., B.S., W.H.T., F.J.M.), Microbiology (J.C.D.H.), Anatomy and Cell Biology (J.S.), Biochemistry (L.G.), and Protein Crystallography Facility (L.G.), University of Iowa, Iowa City; and Veterans Affair Medical Center, Iowa City, IA (F.J.M.)Bojana Stanic - From the Departments of Internal Medicine (B.M.S., S.J., B.S., W.H.T., F.J.M.), Microbiology (J.C.D.H.), Anatomy and Cell Biology (J.S.), Biochemistry (L.G.), and Protein Crystallography Facility (L.G.), University of Iowa, Iowa City; and Veterans Affair Medical Center, Iowa City, IA (F.J.M.)William H Thiel - From the Departments of Internal Medicine (B.M.S., S.J., B.S., W.H.T., F.J.M.), Microbiology (J.C.D.H.), Anatomy and Cell Biology (J.S.), Biochemistry (L.G.), and Protein Crystallography Facility (L.G.), University of Iowa, Iowa City; and Veterans Affair Medical Center, Iowa City, IA (F.J.M.)Lokesh Gakhar - From the Departments of Internal Medicine (B.M.S., S.J., B.S., W.H.T., F.J.M.), Microbiology (J.C.D.H.), Anatomy and Cell Biology (J.S.), Biochemistry (L.G.), and Protein Crystallography Facility (L.G.), University of Iowa, Iowa City; and Veterans Affair Medical Center, Iowa City, IA (F.J.M.)Jon C D Houtman - From the Departments of Internal Medicine (B.M.S., S.J., B.S., W.H.T., F.J.M.), Microbiology (J.C.D.H.), Anatomy and Cell Biology (J.S.), Biochemistry (L.G.), and Protein Crystallography Facility (L.G.), University of Iowa, Iowa City; and Veterans Affair Medical Center, Iowa City, IA (F.J.M.)Francis J Miller Jr - From the Departments of Internal Medicine (B.M.S., S.J., B.S., W.H.T., F.J.M.), Microbiology (J.C.D.H.), Anatomy and Cell Biology (J.S.), Biochemistry (L.G.), and Protein Crystallography Facility (L.G.), University of Iowa, Iowa City; and Veterans Affair Medical Center, Iowa City, IA (F.J.M.). francis-miller@uiowa.edu
- Resource Type
- Journal article
- Publication Details
- Circulation research, Vol.115(11), pp.911-918
- DOI
- 10.1161/CIRCRESAHA.115.304267
- PMID
- 25228390
- PMCID
- PMC5025877
- ISSN
- 0009-7330
- eISSN
- 1524-4571
- Grant note
- R01 HL081750 / NHLBI NIH HHS HL081750 / NHLBI NIH HHS I01 BX001729 / BLRD VA CA136729 / NCI NIH HHS R01 CA136729 / NCI NIH HHS
- Language
- English
- Date published
- 11/07/2014
- Academic Unit
- Microbiology and Immunology; Cardiovascular Medicine; Obstetrics and Gynecology; Biochemistry and Molecular Biology; Medicine Administration; Internal Medicine
- Record Identifier
- 9984094558402771
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