title: Substitution of methionine 63 or 83 in S100A9 and cysteine 42 in S100A8 abrogate the antifungal activities of S100A8/A9: potential role for oxidative regulation.
abstract: S100A8 and S100A9 and their heterocomplex calprotectin (S100A8/A9) are abundant cytosolic constituents in human neutrophils previously shown to possess antifungal activity. This study was designed to investigate mechanisms involved in the modulation of the antifungal properties of S100A8/A9. S100A8, S100A9 and site-directed mutants of both proteins were tested for their antifungal effect against Candida albicans in microplate dilution assays. Whereas S100A8 alone did not inhibit fungal growth, S100A9 by itself had a moderate antifungal effect. Combining both proteins had the strongest effect. Supporting a potential role for oxidation in S100A8/A9, substitution of methionine 63 or 83 of S100A9 resulted in the loss of antifungal activity. Additionally, the substitution to alanine of cysteine 42 of S100A8 also caused a loss of S100A8's ability to enhance S100A9's antifungal effect. Overall, our data indicate that both S100A8 and S100A9 are required for their fully active antifungal effect and that oxidation regulates S100A8/A9 antifungal activity through mechanisms that remain to be elucidated and evaluated. Finally, together with our previous work describing the oxidation-sensitive anti-inflammatory effects of S100A8/A9, we propose that S100A8/A9 exerts an anti-inflammatory activity in healthy state and that conditions associated with oxidative stress activate the antifungal activity of S100A8/A9.
authors.affiliation: Department of Oral Medicine and Diagnostic Sciences, UIC College of Dentistry (M/C 838), University of Illinois, Chicago, IL 60612-7213, USA. sroussih@uic.edu
authors.forename: Herve Y
authors.initials: HY
authors.lastname: Sroussi
authors.forename: Gerwald A
authors.initials: GA
authors.lastname: Köhler
authors.forename: Nina
authors.initials: N
authors.lastname: Agabian
authors.forename: Dana
authors.initials: D
authors.lastname: Villines
authors.forename: Joel M
authors.initials: JM
authors.lastname: Palefsky
mesh_terms: D000935:Antifungal Agents / Q000378:metabolism*; D040501:Calgranulin A / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D040502:Calgranulin B / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002176:Candida albicans / Q000254:growth & development / Q000276:immunology*; D006801:Humans; D008826:Microbial Sensitivity Tests; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000235:genetics; D020125:Mutation, Missense; D010084:Oxidation-Reduction
chemical_list: D000935:Antifungal Agents; D040501:Calgranulin A; D040502:Calgranulin B; D050505:Mutant Proteins