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The FASEB Journal, Vol 2, 2487-2491, Copyright © 1988 by The Federation of American Societies for Experimental Biology

RESEARCH COMMUNICATIONS

Fluorescence-based assay for reactive oxygen species: a protective role for creatinine

AN Glazer
Department of Microbiology and Immunology, University of California, Berkeley 94720.

Attack by reactive oxygen species leads to a decay in phycoerythrin fluorescence emission. This phenomenon provides a versatile new assay for small molecules and macromolecules that can function as protective compounds. With 1-2 x 10(-8) M phycoerythrin, under conditions where peroxyl radical generation is rate-limiting, the fluorescence decay follows apparent zero-order kinetics. On reaction with HO., generated with the ascorbate-Cu2+ system, the fluorescence decays with apparent first-order kinetics. Examination of the major components of human urine in this assay confirms that at physiological concentrations, urate protects against both types of oxygen radicals. A novel finding is that creatinine protects efficiently by a chelation mechanism against radical damage in the ascorbate-Cu2+ system at creatinine, ascorbate, and Cu2+ concentrations comparable to those in normal urine. Urate and creatinine provide complementary modes of protection against reactive oxygen species in the urinary tract.


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Copyright © 1988 by The Federation of American Societies for Experimental Biology.