The NADPH oxidase 2 subunit p47phox binds to the WAVE regulatory complex and p22phox in a mutually exclusive manner

J Biol Chem. 2024 Apr;300(4):107130. doi: 10.1016/j.jbc.2024.107130. Epub 2024 Mar 1.

Abstract

The actin cytoskeleton and reactive oxygen species (ROS) both play crucial roles in various cellular processes. Previous research indicated a direct interaction between two key components of these systems: the WAVE1 subunit of the WAVE regulatory complex (WRC), which promotes actin polymerization and the p47phox subunit of the NADPH oxidase 2 complex (NOX2), which produces ROS. Here, using carefully characterized recombinant proteins, we find that activated p47phox uses its dual Src homology 3 domains to bind to multiple regions within the WAVE1 and Abi2 subunits of the WRC, without altering WRC's activity in promoting Arp2/3-mediated actin polymerization. Notably, contrary to previous findings, p47phox uses the same binding pocket to interact with both the WRC and the p22phox subunit of NOX2, albeit in a mutually exclusive manner. This observation suggests that when activated, p47phox may separately participate in two distinct processes: assembling into NOX2 to promote ROS production and engaging with WRC to regulate the actin cytoskeleton.

Keywords: Arp2/3; NADPH oxidase; NOX2; ROS; WAVE complex; WRC; actin cytoskeleton; p47phox.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / genetics
  • Actin Cytoskeleton / metabolism
  • Actin-Related Protein 2-3 Complex / metabolism
  • Actins / metabolism
  • Binding Sites
  • Humans
  • NADPH Oxidase 2* / genetics
  • NADPH Oxidase 2* / metabolism
  • NADPH Oxidases / genetics
  • NADPH Oxidases / metabolism
  • Protein Binding
  • Protein Subunits / genetics
  • Protein Subunits / metabolism
  • Reactive Oxygen Species / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Wiskott-Aldrich Syndrome Protein Family* / genetics
  • Wiskott-Aldrich Syndrome Protein Family* / metabolism

Substances

  • Actins
  • CYBA protein, human
  • CYBB protein, human
  • NADPH Oxidase 2
  • NADPH Oxidases
  • neutrophil cytosolic factor 1
  • Reactive Oxygen Species
  • Wiskott-Aldrich Syndrome Protein Family
  • Recombinant Proteins
  • Protein Subunits
  • Actin-Related Protein 2-3 Complex