Assembly and activation of the phagocyte NADPH oxidase

Specific interaction of the N-terminal Src homology 3 domain of p47(phox) with p22(phox) is required for activation of the NADPH oxidase

Hideki Sumimoto, Kenichiro Hata, Kazuhito Mizuki, Takashi Ito, Yohko Kage, Yoshiyuki Sakaki, Yasuyuki Fukumaki, Michio Nakamura, Koichiro Takeshige

Research output: Contribution to journalArticle

151 Citations (Scopus)

Abstract

The phagocyte NADPH oxidase is activated during phagocytosis to produce superoxide, a precursor of microbicidal oxidants. The activation involves assembly of membrane-integrated cytochrome b558 comprising gp91(phox) and p22(phox), two specialized cytosolic proteins (p47(phox) and p67(phox)), each containing two Src homology 3 (SH3) domains, and the small G protein Rac. In the present study, we show that the N-terminal SH3 domain of p47(phox) binds to the C-terminal cytoplasmic tail of p22(phox) with high affinity (K(D) = 0.34 μM). The binding is specific to this domain among several SH3 domains including the C-terminal one of p47(phox) and the two of p67(phox) and requires the Pro156-containing proline-rich sequence but not other putative SH3 domain-binding sites of p22(phox). Replacement of Trp193 by Arg in the N-terminal SH3 domain completely abrogates the association with p22(phox). A mutant p47(phox) with this substitution is incapable of supporting superoxide production under cell-free activation conditions. These findings provide direct evidence that the interaction between the N-terminal SH3 domain of p47(phox) and the proline-rich region of p22(phox) is essential for activation of the NADPH oxidase.

Original languageEnglish
Pages (from-to)22152-22158
Number of pages7
JournalJournal of Biological Chemistry
Volume271
Issue number36
DOIs
Publication statusPublished - Sep 25 1996

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src Homology Domains
NADPH Oxidase
Phagocytes
Chemical activation
Proline
Superoxides
4-ethoxymethylene-2-phenyl-2-oxazoline-5-one
Monomeric GTP-Binding Proteins
Oxidants
Phagocytosis
Substitution reactions
Binding Sites
Association reactions
Membranes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Assembly and activation of the phagocyte NADPH oxidase : Specific interaction of the N-terminal Src homology 3 domain of p47(phox) with p22(phox) is required for activation of the NADPH oxidase. / Sumimoto, Hideki; Hata, Kenichiro; Mizuki, Kazuhito; Ito, Takashi; Kage, Yohko; Sakaki, Yoshiyuki; Fukumaki, Yasuyuki; Nakamura, Michio; Takeshige, Koichiro.

In: Journal of Biological Chemistry, Vol. 271, No. 36, 25.09.1996, p. 22152-22158.

Research output: Contribution to journalArticle

Sumimoto, Hideki ; Hata, Kenichiro ; Mizuki, Kazuhito ; Ito, Takashi ; Kage, Yohko ; Sakaki, Yoshiyuki ; Fukumaki, Yasuyuki ; Nakamura, Michio ; Takeshige, Koichiro. / Assembly and activation of the phagocyte NADPH oxidase : Specific interaction of the N-terminal Src homology 3 domain of p47(phox) with p22(phox) is required for activation of the NADPH oxidase. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 36. pp. 22152-22158.
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