Journal article
Critical roles for p22phox in the structural maturation and subcellular targeting of Nox3
Biochemical journal, Vol.403(1), pp.97-108
04/01/2007
DOI: 10.1042/BJ20060819
PMCID: PMC1828898
PMID: 17140397
Abstract
Otoconia are small biominerals in the inner ear that are indispensable for the normal perception of gravity and motion. Normal otoconia biogenesis requires Nox3, a Nox (NADPH oxidase) highly expressed in the vestibular system. In HEK-293 cells (human embryonic kidney cells) transfected with the Nox regulatory subunits NoxO1 (Nox organizer 1) and NoxA1 (Nox activator 1), functional murine Nox3 was expressed in the plasma membrane and exhibited a haem spectrum identical with that of Nox2, the electron transferase of the phagocyte Nox. In vitro Nox3 cDNA expressed an approximately 50 kDa primary translation product that underwent N-linked glycosylation in the presence of canine microsomes. RNAi (RNA interference)-mediated reduction of endogenous p22phox, a subunit essential for stabilization of Nox2 in phagocytes, decreased Nox3 activity in reconstituted HEK-293 cells. p22phox co-precipitated not only with Nox3 and NoxO1 from transfectants expressing all three proteins, but also with NoxO1 in the absence of Nox3, indicating that p22phox physically associated with both Nox3 and with NoxO1. The plasma membrane localization of Nox3 but not of NoxO1 required p22phox. Moreover, the glycosylation and maturation of Nox3 required p22phox expression, suggesting that p22phox was required for the proper biosynthesis and function of Nox3. Taken together, these studies demonstrate critical roles for p22phox at several distinct points in the maturation and assembly of a functionally competent Nox3 in the plasma membrane.
Details
- Title: Subtitle
- Critical roles for p22phox in the structural maturation and subcellular targeting of Nox3
- Creators
- Yoko Nakano - University of IowaBotond Banfi - University of IowaAlgirdas J Jesaitis - Montana State UniversityMary C Dinauer - Indiana UniversityLee-Ann H Allen - University of IowaWilliam M Nauseef - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Biochemical journal, Vol.403(1), pp.97-108
- DOI
- 10.1042/BJ20060819
- PMID
- 17140397
- PMCID
- PMC1828898
- ISSN
- 0264-6021
- eISSN
- 1470-8728
- Grant note
- I01 BX000513 / BLRD VA R01 HL053592 / NHLBI NIH HHS R01 AI026711 / NIAID NIH HHS R01 HL045635 / NHLBI NIH HHS HL53592 / NHLBI NIH HHS AI34879 / NIAID NIH HHS R01 AI034879 / NIAID NIH HHS R01 AI26711 / NIAID NIH HHS R56 AI034879 / NIAID NIH HHS
- Language
- English
- Date published
- 04/01/2007
- Academic Unit
- Microbiology and Immunology; Anatomy and Cell Biology; Infectious Diseases; Otolaryngology; Internal Medicine
- Record Identifier
- 9984284355702771
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