Spermine suppresses the activation of human neutrophil NADPH oxidase in cell-free and semi-recombinant systems.
Spermine, a cellular polyamine, down-regulates O2- generation in human neutrophils stimulated by receptor-linked agonist [Ogata, Tamura and Takeshita (1992) Biochem. Biophys. Res. Commun. 182, 20-26]. In this study, to elucidate the mechanism for the inhibition, the effect of spermine on cell-free activation of the O2- generating enzyme (NADPH oxidase) was examined. Spermine suppressed the SDS-induced activation of NADPH oxidase in a dose-dependent manner with an IC50 of 18 microM. The inhibition was specific for spermine over its precursor amines, spermidine and putrescine. Spermine did not alter the Km for NADPH or the optimal concentration of SDS for activation. The amine was inhibitory only when added before activation, indicating that it affects the activation process rather than the enzyme's activity. An increased concentration of cytosol partly prevented the inhibition by spermine. In semi-recombinant cell-free system, spermine inhibited the activation of NADPH oxidase as effectively as in the cell-free system (IC50 = 13 microM). Pretreatment of each recombinant cytosolic component with spermine revealed that they (especially p67phox) are sensitive to spermine. These results suggest that spermine interacts with cytosolic component(s) and impairs the assembly of NADPH oxidase.
Ogata
K
K
Department of Biochemistry, Oita Medical University, Japan.
Nishimoto
N
N
Uhlinger
D J
DJ
Igarashi
K
K
Takeshita
M
M
Tamura
M
M
D002474:Cell-Free System; D003600:Cytosol / Q000378:metabolism; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D009247:NADH, NADPH Oxidoreductases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D019255:NADPH Oxidases; D009504:Neutrophils / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D013096:Spermine / Q000494:pharmacology*
D011994:Recombinant Proteins; D013096:Spermine; D012967:Sodium Dodecyl Sulfate; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases