Identification of an actin-binding site in p47phox an organizer protein of NADPH oxidase.
Actin has been reported to enhance the superoxide-generating activity of neutrophil NADPH oxidase in a cell-free system and to interact with p47phox, a regulatory subunit of the oxidase. In the present study, we searched for an actin-binding site in p47phox by far-western blotting and blot-binding assays using truncated forms of ... p47phox. The amino-acid sequence 319-337 was identified as an actin-binding site, and a synthetic peptide of this sequence bound to actin. The sequence shows no homology to other known actin-binding motifs. It is located in the autoinhibitory region of p47phox and includes Ser-328, a phosphorylation site essential for unmasking. Although a phosphorylation-mimetic p47phox mutant bound to actin with a lower affinity than the wild type, the same mutant interacted with filamentous actin more efficiently than the wild type. A mutant peptide p47phox (319-337, Ser328Glu) bound to filamentous actin more tightly than to monomer actin. These results suggest that p47phox moves to cortical actin when it becomes unmasked in the cells.
Mesh Terms:
Actin Cytoskeleton, Actins, Amino Acid Motifs, Amino Acid Sequence, Amino Acid Substitution, Animals, Binding Sites, Cell Line, Cell-Free System, Humans, NADPH Oxidase, Peptides, Phosphoproteins, Point Mutation, Protein Binding
Actin Cytoskeleton, Actins, Amino Acid Motifs, Amino Acid Sequence, Amino Acid Substitution, Animals, Binding Sites, Cell Line, Cell-Free System, Humans, NADPH Oxidase, Peptides, Phosphoproteins, Point Mutation, Protein Binding
FEBS Lett.
Date: Jan. 09, 2006
PubMed ID: 16375898
View in: Pubmed Google Scholar
Download Curated Data For This Publication
151794
Switch View:
- Interactions 2