Effect of glutaraldehyde on NADPH oxidase system of guinea pig polymorphonuclear leukocytes.
In an attempt to elucidate properties and activation mechanisms of the NADPH oxidase system, which is known to be responsible for the production of superoxide anion (O2-) in cell membranes of polymorphonuclear leukocytes (PMNL), intact guinea pig PMNL were treated with glutaraldehyde, a protein crosslinking reagent, before or after stimulation with phorbol 12-myristate 13-acetate (PMA). Then, PMNL were disrupted and NADPH oxidase activity was measured. After the treatment of resting PMNL with glutaraldehyde, NADPH oxidase was no longer activated by PMA. On the other hand, the NADPH oxidase activity enhanced by PMA in advance was markedly retained by the glutaraldehyde treatment of such PMA-stimulated PMNL as compared to that in untreated cells. Similar retention by glutaraldehyde of the stimulated NADPH oxidase activity was observed in PMNL stimulated by formyl-methionyl-leucyl-phenylalanine (FMLP) and cytochalasin D. Furthermore, the oxidase activity of glutaraldehyde-treated PMNL was stable during incubation at 37 degrees C, the half life of the oxidase activity of the treated PMNL being more than 90 min whereas that of the untreated PMNL is about 15 min. This ability of the glutaraldehyde treatment to retain the activity was also observed against inactivation by high concentrations of NaCl and by positively charged alkylamine.
Katayama
T
T
Department of Physiological Chemistry, Hiroshima University School of Medicine, Japan.
Ohtsuka
T
T
Wakamura
K
K
Yoshida
K
K
Okamura
N
N
Ishibashi
S
S
D000447:Aldehydes / Q000494:pharmacology*; D000588:Amines / Q000494:pharmacology; D000818:Animals; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D005976:Glutaral / Q000494:pharmacology*; D006168:Guinea Pigs; D006358:Hot Temperature; D006838:Hydrocarbons; D007700:Kinetics; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D019255:NADPH Oxidases; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology*; D012965:Sodium Chloride / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism
D000447:Aldehydes; D000588:Amines; D006838:Hydrocarbons; D013481:Superoxides; D012965:Sodium Chloride; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; C013553:hexadecylamine; D005976:Glutaral