The cell-free activation of human neutrophil NADPH oxidase (O2--generating enzyme) is enhanced by actin [Morimatsu, Kawagoshi, Yoshida and Tamura (1997) Biochem. Biophys. Res. Commun. 230, 206-210]. In an attempt to elucidate the mechanism, we examined the effect of actin-depolymerizing agents on the duration of NADPH oxidase in a cell-free system. The addition of DNase I, an F-actin-depolymerizing protein, caused an accelerated deactivation of the oxidase. The deactivation was also facilitated by latrunculin A, a sponge toxin that depolymerizes F-actin. Exogenously added actin prevented the deactivation by DNase I or latrunculin A, whereas EDTA accelerated a dilution-induced deactivation of the oxidase and Mg2+ ions retarded it. The stability in dilution was found to correlate well with free Mg2+ concentration. Estimation of F-actin in the system showed that F-actin increased during the oxidase activation and that DNase I or EDTA decreased F-actin content in parallel with the activity. Treatment of the cell-free mixture with a chemical cross-linker prevented the deactivation and F-actin decrease by EDTA. Taken together, these results suggest that actin filaments which grow during the activation of NADPH oxidase prolong the lifetime of the oxidase.
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Research Article|
June 26 2000
Deactivation of neutrophil NADPH oxidase by actin-depolymerizing agents in a cell-free system
Minoru TAMURA;
Minoru TAMURA
1
1To whom correspondence should be addressed (e-mail miketamu@;en3.ehime-u.ac.jp).
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Masamichi KANNO;
Masamichi KANNO
1Department of Applied Biochemistry, Faculty of Engineering, Ehime University, Matsuyama, Ehime 790-8577, Japan
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Yaeta ENDO
Yaeta ENDO
1Department of Applied Biochemistry, Faculty of Engineering, Ehime University, Matsuyama, Ehime 790-8577, Japan
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Publisher: Portland Press Ltd
Received:
November 29 1999
Revision Received:
March 20 2000
Accepted:
April 16 2000
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 2000
2000
Biochem J (2000) 349 (1): 369–375.
Article history
Received:
November 29 1999
Revision Received:
March 20 2000
Accepted:
April 16 2000
Citation
Minoru TAMURA, Masamichi KANNO, Yaeta ENDO; Deactivation of neutrophil NADPH oxidase by actin-depolymerizing agents in a cell-free system. Biochem J 1 July 2000; 349 (1): 369–375. doi: https://doi.org/10.1042/bj3490369
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