Fungal metabolite gliotoxin inhibits assembly of the human respiratory burst NADPH oxidase

Abstract

ABSTRACTReactive oxygen species are a critical weapon in the killing ofAspergillus fumigatusby polymorphonuclear leukocytes (PMN), as demonstrated by severe aspergillosis in chronic granulomatous disease. In the present study,A. fumigatus-produced mycotoxins (fumagillin, gliotoxin [GT], and helvolic acid) are examined for their effects on the NADPH oxidase activity in human PMN. Of these mycotoxins, only GT significantly and stoichiometrically inhibits phorbol myristate acetate (PMA)-stimulated O2−generation, while the other two toxins are ineffective. The inhibition is dependent on the disulfide bridge of GT, which interferes with oxidase activation but not catalysis of the activated oxidase. Specifically, GT inhibits PMA-stimulated events: p47phoxphosphorylation, its incorporation into the cytoskeleton, and the membrane translocation of p67phox, p47phox, and p40phox, which are crucial steps in the assembly of the active NADPH oxidase. Thus, damage to p47phoxphosphorylation is likely a key to inhibiting NADPH oxidase activation. GT does not inhibit the membrane translocation of Rac2. The inhibition of p47phoxphosphorylation is due to the defective membrane translocation of protein kinase C (PKC) βII rather than an effect of GT on PKC βII activity, suggesting a failure of PKC βII to associate with the substrate, p47phox, on the membrane. These results suggest thatA. fumigatusmay confront PMN by inhibiting the assembly of the NADPH oxidase with its hyphal product, GT.