Salicylic acid-induced inactivation of creatine kinase in the presence of lactoperoxidase and H2O2.
To clarify one mechanism of aspirin-induced gastric mucosal damage, inactivation of creatine kinase (CK) by salicylic acid that is easily produced from aspirin in vivo was examined in the presence of lactoperoxidase (LPO) and H2O2 (LPO-H2O2). Salicylic acid inactivated CK (rabbit muscle) during its interaction with LPO-H2O2. CK activity in gastric mucosal homogenate decreased dependent on the concentration of salicylic acid in the presence of LPO-H2O2. Oxygen radical scavengers did not prevent the inactivation of CK. Direct detection of free radicals of salicylic acid by electron spin resonance was unsuccessful. However, glutathionyl radicals were formed during the interaction of salicylic acid with LPO-H2O2 in the presence of reduced glutathione and 5,5-dimethyl-1-pyrroline oxide as a spin trap agent. Among salicylic acid-related drugs, salsalate, but not aspirin and ethenzamide, inactivated CK, indicating the phenolic hydroxyl group is oxidized by LPO-H2O2. During oxidation of salicylic acid by LPO-H2O2, the sulfhydryl group in CK markedly decreased, and salicylic acid bound to CK. These results indicate that CK was inactivated through loss of the sulfhydryl group and binding of salicylic acid.
Muraoka
Sanae
S
Hokkaido College of Pharmacy, Department of Biochemistry, Katuraoka-cho 7-1, Otaru 047-0264, Japan.
Miura
Toshiaki
T
D000818:Animals; D003402:Creatine Kinase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D016166:Free Radical Scavengers / Q000494:pharmacology; D005753:Gastric Mucosa / Q000201:enzymology*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D066298:In Vitro Techniques; D007784:Lactoperoxidase / Q000494:pharmacology; D008297:Male; D010084:Oxidation-Reduction; D051381:Rats; D017208:Rats, Wistar; D020156:Salicylic Acid / Q000494:pharmacology*; D013438:Sulfhydryl Compounds / Q000032:analysis / Q000378:metabolism
D016166:Free Radical Scavengers; D013438:Sulfhydryl Compounds; D006861:Hydrogen Peroxide; D007784:Lactoperoxidase; D003402:Creatine Kinase; D020156:Salicylic Acid