Noxa1 as a moderate activator of Nox2-based NADPH oxidase

Masahito Kawano, Kazuhiro Miyamoto, Yuki Kaito, Hideki Sumimoto, Minoru Tamura

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Abstract

Noxa1 was discovered as an activating factor for Nox1, an O2 - generating enzyme. Subsequent studies have shown that Noxa1 is colocalized with Nox2 in several cell types, including vascular cells. Nox2 activation by Noxa1 has been examined in reconstituted model cells. However, little is known about the kinetic properties of Noxa1 in Nox2 activation. In the present study, we used purified cyt.b 558 (Nox2 plus p22 phox), Rac(Q61L), and Noxo1 to examine the ability of Noxa1 to activate Nox2. In the pure reconstitution system, Noxa1 activated Nox2 with lower efficiency than p67 phox, a canonical activator of Nox2. The EC 50 value of Noxa1 was considerably higher than that of p67 phox. The V max value with Noxa1 and Noxo1 was one-third of that with p67 phox and p47 phox. The EC 50 value of Noxo1 or Rac(Q61L) was also higher when Noxa1 was used. The affinity of FAD for the oxidase and the stability of the active complex were remarkably low when Noxa1 and Noxo1 were used compared with p67 phox and p47 phox. The stability was not improved by fusion of Noxa1 with Rac(Q61L). These findings show that Noxa1 has quite different kinetic properties from p67 phox and suggest that Noxa1 may function as a moderate activator of Nox2.

Original languageEnglish
Pages (from-to)1-7
Number of pages7
JournalArchives of Biochemistry and Biophysics
Volume519
Issue number1
DOIs
Publication statusPublished - Mar 1 2012

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NADPH Oxidase
Chemical activation
Flavin-Adenine Dinucleotide
Kinetics
Blood Vessels
Oxidoreductases
Fusion reactions
neutrophil cytosol factor 67K
Enzymes
4-ethoxymethylene-2-phenyl-2-oxazoline-5-one

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Noxa1 as a moderate activator of Nox2-based NADPH oxidase. / Kawano, Masahito; Miyamoto, Kazuhiro; Kaito, Yuki; Sumimoto, Hideki; Tamura, Minoru.

In: Archives of Biochemistry and Biophysics, Vol. 519, No. 1, 01.03.2012, p. 1-7.

Research output: Contribution to journalArticle

Kawano, Masahito ; Miyamoto, Kazuhiro ; Kaito, Yuki ; Sumimoto, Hideki ; Tamura, Minoru. / Noxa1 as a moderate activator of Nox2-based NADPH oxidase. In: Archives of Biochemistry and Biophysics. 2012 ; Vol. 519, No. 1. pp. 1-7.
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