Oxidation of sulfhydryl groups and inhibition of the (Ca2+ + Mg2+)-ATPase by arsenazo III.
In the presence of divalent cations, the metallochromic Ca2+ indicator arsenazo III is reduced by sulfhydryl groups to form an azo anion radical. Reduced arsenazo III is reoxidized back to its original state by oxygen. The formation of the arsenazo III azo anion radical in the presence of sarcoplasmic reticulum vesicles leads to the rapid inhibition of the (Ca2+ + Mg2+)-ATPase. These data indicate that several factors should be considered when arsenazo III is used as a Ca2+ indicator; (1) Functionally important sulfhydryl groups may be oxidized by arsenazo III; (2) the generation of free radicals by arsenazo III reduction may be toxic to the system being studied; (3) the absorbance spectrum of arsenazo III is altered when reduced by sulfhydryl groups.
Beeler
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Department of Biochemistry, Uniformed Services University of the Health Sciences, Bethesda, MD 20814.
D000818:Animals; D001150:Arsenazo III / Q000494:pharmacology*; D001391:Azo Compounds / Q000494:pharmacology*; D017301:Ca(2+) Mg(2+)-ATPase / Q000037:antagonists & inhibitors*; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors*; D004789:Enzyme Activation / Q000187:drug effects; D005609:Free Radicals; D007700:Kinetics; D010084:Oxidation-Reduction; D051381:Rats; D011919:Rats, Inbred Strains; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000201:enzymology*; D013438:Sulfhydryl Compounds / Q000378:metabolism*
D001391:Azo Compounds; D005609:Free Radicals; D013438:Sulfhydryl Compounds; D001150:Arsenazo III; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases