Uric acid mediates photodynamic inactivation of caprine alpha-2-macroglobulin.
Uric acid (2,6,8 trioxopurine), the end product of purine metabolism in mammalian systems, has shown a wide range of antioxidant properties including scavenging of hydroxyl radical and singlet oxygen. In this study we show that in the presence of visible light, uric acid disrupted caprine alpha-2-macroglobulin (alpha(2) M) structure and antiproteolytic function in vitro. Proteinase cleaves the bait region of caprine inhibitor inducing major conformational changes and entrapping the enzyme within its molecular cage. In contrast to native alpha(2) M, modified antiproteinase lost half of its antiproteolytic potential within 4 hours of uric acid exposure. The changes in uv-absorption spectra of the treated protein suggested possible spatial rearrangement of subunits or conformational change. Analysis of the mechanism by which alpha(2) M was inactivated revealed that the process was dependent on generation of superoxide anion and hydrogen peroxide. Our findings suggest that antiproteolytic activity of caprine alpha(2) M could be compromised via oxidative modification mediated by uric acid. Moreover, low concentrations of alpha(2) M were found to stimulate superoxide production by some unknown mechanism.
Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh - 202 002, INDIA.
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D020816:Amino Acid Motifs; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D006041:Goats; D006861:Hydrogen Peroxide / Q000378:metabolism; D008027:Light; D010084:Oxidation-Reduction; D010777:Photochemistry; D011487:Protein Conformation / Q000187:drug effects; D013056:Spectrophotometry, Ultraviolet; D013481:Superoxides / Q000378:metabolism; D014527:Uric Acid / Q000494:pharmacology*; D000511:alpha-Macroglobulins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000494:pharmacology / Q000528:radiation effects
D000511:alpha-Macroglobulins; D013481:Superoxides; D014527:Uric Acid; D006861:Hydrogen Peroxide