Phycoerythrin fluorescence-based assay for peroxy radicals: A screen for biologically relevant protective agents

doi:10.1016/0003-2697(89)90056-0

Analytical Biochemistry

Volume 177, Issue 2, March 1989, Pages 300-306

https://doi.org/10.1016/0003-2697(89)90056-0 Get rights and content

Abstract

Under the conditions of this assay, antioxidants that react rapidly with peroxy free radicals (e.g., ascorbate, vitamin E analogs, urate), protect phycoerythrin completely from damage by such radicals generated by thermal decomposition of 2,2′-azobis(2-amidinopropane); other compounds provide partial concentration-dependent protection. Change in phycoerythrin fluorescence emission with time provides a measure of the rate of free radical damage. The assay exploits the unusual reactivity of phycoerythrin toward these peroxy radicals. On a molar basis, phycoerythrin reacts with these radicals over 100-fold slower than do ascorbate or vitamin E analogs, but over 60-fold faster than other proteins. Applications of this assay to the estimation of the peroxy radical scavenging capacity of human plasma are described, and to the comparison of the scavenging properties of several proteins and of DNA, of vitamins and their derivatives, of catecholamine neurotransmitters, and of a variety of other low molecular weight biological compounds.

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^☆: This work was supported by National Institutes of Health Grant GM 28994 and by National Science Foundation Grant DMB 8518066 to A.N.G.

²: On sabbatical leave from the Department of Biological Chemistry, UCLA School of Medicine, Los Angeles, CA 90024.

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Phycoerythrin fluorescence-based assay for peroxy radicals: A screen for biologically relevant protective agents☆

Abstract

Biochim. Biophys. Acta

J. Biol. Chem

J. Biol. Chem

Anal. Biochem

J. Free Radicals Biol. Med

FEBS Lett

Arch. Biochem. Biophys

Trends Biochem. Sci

J. Lipids Res

Arch. Biochem. Biophys

Biochem. Biophys. Res. Commun

Biochim. Biophys. Acta