Oxidation of methionine 63 and 83 regulates the effect of S100A9 on the migration of neutrophils in vitro.
The calcium-binding proteins S100A8 and S100A9 and their heterocomplex calprotectin are abundant cytosolic constituents in human neutrophils, constitutively expressed by mucosal epithelium and in association with inflammation by epidermal keratinocytes. S100A8 and S100A9 are pleiotropic proteins, which partake in the regulation of leukocyte migration. This study was designed to investigate the effect of S100A9 on neutrophil migration and to explore the mechanisms that regulate this effect. Based on previous results with S100A8, we hypothesized that S100A9 repels neutrophils and that oxidation of S100A9 regulates this function. Using standard Transwell chemotaxis assays and site-directed mutagenesis, we show that S100A9 exerts a chemo-repulsive (fugetactic) effect on peripheral neutrophils, an effect abolished by oxidation of S100A9. After substitution of methionine 63 and 83 for alanine, S100A9 maintained its fugetaxis activity, even in inhibitory, oxidative conditions. Together, the data suggest that S100A9 serves as a molecular switch for oxidative control of inflammation regulated by the oxidation of species-conserved methionine residues. In healthy mucosal tissue, expression of S100A9 by the epithelium may serve to inhibit leukocyte recruitment. However, conditions of oxidative stress, including infection and overgrowth of opportunistic pathogens, may abrogate this activity by neutralizing S100A9 as a result of its oxidative alteration.
Sroussi
Herve Y
HY
Department of Oral Medicine, College of Dentistry, University of Illinois at Chicago, Chicago, IL 60612-7213, USA. sroussih@uic.edu
Berline
Jennifer
J
Palefsky
Joel M
JM
D000818:Animals; D040502:Calgranulin B / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D002465:Cell Movement / Q000187:drug effects / Q000276:immunology; D002633:Chemotaxis / Q000187:drug effects / Q000276:immunology; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D008715:Methionine / Q000378:metabolism*; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology*; D010084:Oxidation-Reduction; D017354:Point Mutation; D051381:Rats
D040502:Calgranulin B; D009240:N-Formylmethionine Leucyl-Phenylalanine; D008715:Methionine