Inactivation of NADPH oxidase from human neutrophils by affinity labeling with pyridoxal 5'-diphospho-5'-adenosine.
When a particulate NADPH oxidase prepared from phorbol ester-activated human neutrophils was treated with pyridoxal 5'-diphospho-5'-adenosine (PLP-AMP), the superoxide anion-producing activity was inhibited according to affinity labeling kinetics. NADPH afforded a protection against inactivation which was competitive with respect to PLP-AMP; 2',5'-ADP and 2'-phospho-5' diphosphoadenosine (ATP ribose) appeared to be as potent as NADPH as protecting agents. NADP+ and ATP were less effective, while ADP and GTP-gamma-S did not protect significantly. These results suggest that PLP-AMP can be used, in conjunction with tritiated cyanoborohydride, to identify the elusive NADPH-dependent flavoprotein which is part of the electron transfer chain of NADPH oxidase.
Ravel
P
P
CNRS URA 1461, Hôpital Necker, Paris, France.
Lederer
F
F
D000244:Adenosine Diphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D000345:Affinity Labels / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008433:Mathematics; D009247:NADH, NADPH Oxidoreductases / Q000037:antagonists & inhibitors* / Q000097:blood; D009249:NADP / Q000031:analogs & derivatives / Q000494:pharmacology*; D019255:NADPH Oxidases; D009504:Neutrophils / Q000201:enzymology*; D010084:Oxidation-Reduction; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives*; D013329:Structure-Activity Relationship
D000345:Affinity Labels; D009249:NADP; D011732:Pyridoxal Phosphate; D000244:Adenosine Diphosphate; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; C048236:pyridoxal 5'-diphospho-5'-adenosine