Microsomal UDP-glucuronyltransferase in rat liver: Oxidative activation

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Microsomal UDP-glucuronyltransferase in rat liver: Oxidative activation

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Microsomal UDP-glucuronyltransferase in rat liver: Oxidative activation

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Título: Microsomal UDP-glucuronyltransferase in rat liver: Oxidative activation
Autor: Letelier, María Eugenia; Pimentel Miranda, Alejandra Patricia; Pino, Paulina; Lepe Agrodnik, Ana María; Faúndez Cáceres, Mario Antonio; Aracena Parks, Paula; Speisky C., Hernán
Resumen: Activation of microsomal UDP-glucuronyltransferase (UDPGT) activity by treatment of hepatic microsomes with either detergents or Fe3+/ascorbate pro-oxidant system has been reported; however, definite mechanisms underlying these effects have not been clarified. In this work, we characterize Fe3+/ascorbate-induced activation of UDPGT activity prior to solubilization with Triton X-100 and after the oxidation process provoked the solubilization of the enzyme. We observed a time-dependent increase in UDPGT activity up to 20 min. incubation of the microsomes with Fe3+/ascorbate (3-times); after 20 min. incubation, however, we observed a time-dependent decrease in this activity to basal levels after 4 hr incubation. Treatment of microsomes with 0.1% Triton X-100 (5 min.) lead to a similar increase in UDPGT activity; higher detergent concentrations produced a dose-dependent decrease in this activity to basal levels with 1% Triton X-100. Interestingly, UDPGT activity was susceptible to activation only when associated to microsomal membranes and the loss of activation correlated with the solubilization of this activity. UDPGT activation by either Fe"/ascorbate or Triton X-100 was correlated with an increase in p-nitrophenol apparent K-m and V-max values. This activation was prevented or reversed by the reducing agents glutathione, cysteine or dithiothreitol when it was induced by the Fe3+/ascorbate. Furthermore, the latter provoked a significant decrease in microsomal thiol content, effect not observed after treatment with Triton X-100. Our results suggest that the main mechanism responsible for Fe3+/ascorbate-induced UDPGT activation is likely to be the promotion of protein sulfhydryl oxidation-, this mechanism appears to be different from detergent-induced UDPGT activation.
URI: http://www.captura.uchile.cl/handle/2250/6289
Fecha: 2005-06
Cita del item: BASIC & CLINICAL PHARMACOLOGY & TOXICOLOGY 96(6):480-486


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