Phenylbutazone radicals inactivate creatine kinase.
Creatine kinase (CK) was used as a marker molecule to examine the side effect of damage to tissues by phenylbutazone (PB), an effective drug to treat rheumatic and arthritic diseases, with horseradish peroxidase and hydrogen peroxide (HRP-H(2)O2). PB inactivated CK during its interaction with HRP-H(2) O(2), and inactivated CK in rat heart homogenate. PB carbon-centered radicals were formed during the interaction of PB with HRP-H(2)O2. The CK efficiently reduced electron spin resonance signals of the PB carbon-centered radicals. The spin trap agent 2-methyl-2-nitrosopropane strongly prevented CK inactivation. These results show that CK was inactivated through interaction with PB carbon-centered radicals. Sulfhydryl groups and tryptophan residues in CK were lost during the interaction of PB with HRP-H(2)O2, suggesting that cysteine and tryptophan residues are oxidized by PB carbon-centered radicals. Other enzymes, including alcohol dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, but not lactate dehydrogenase, were also inactivated. Sulfhydryl enzymes seem to be sensitive to attack by PB carbon-centered radicals. Inhibition of SH enzymes may explain some of the deleterious effects induced by PB.
Miura
T
T
Department of Biochemistry, Hokkaido College of Pharmacy Katuraoka-cho 7-1, Otaru 047-02, Japan. miurat@hokuyakudai.ac.jp
Muraoka
S
S
Fujimoto
Y
Y
D000426:Alcohol Dehydrogenase / Q000037:antagonists & inhibitors; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000009:adverse effects / Q000494:pharmacology*; D015415:Biomarkers; D003402:Creatine Kinase / Q000037:antagonists & inhibitors*; D003545:Cysteine / Q000378:metabolism; D005609:Free Radicals; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000037:antagonists & inhibitors; D006735:Horseradish Peroxidase / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000037:antagonists & inhibitors; D008297:Male; D009206:Myocardium / Q000201:enzymology; D009603:Nitroso Compounds / Q000494:pharmacology; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010544:Peroxidases / Q000378:metabolism; D010653:Phenylbutazone / Q000009:adverse effects / Q000494:pharmacology*; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D013113:Spin Labels; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D013552:Swine; D014364:Tryptophan / Q000378:metabolism
D000894:Anti-Inflammatory Agents, Non-Steroidal; D015415:Biomarkers; D005609:Free Radicals; D009603:Nitroso Compounds; D016877:Oxidants; D013113:Spin Labels; D013438:Sulfhydryl Compounds; D014364:Tryptophan; D006861:Hydrogen Peroxide; D000426:Alcohol Dehydrogenase; D007770:L-Lactate Dehydrogenase; D006735:Horseradish Peroxidase; D010544:Peroxidases; D011451:Prostaglandin-Endoperoxide Synthases; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D003402:Creatine Kinase; D010653:Phenylbutazone; C034526:tert-nitrosobutane; D003545:Cysteine