Purification of mouse liver UDPglucuronosyltransferase.
The present study describes a method for the purification of hepatic microsomal UDPglucuronosyltransferase from two different strains of mice, by a combination of detergent solubilization, ion-exchange chromatography and affinity chromatography. Using this procedure up to 0.5 mg of UDPglucuronosyltransferase could be obtained from 30 g of mouse liver and this enzyme could be purified from as little as 10 g of mouse liver. The molecular weight of the apparently homogeneous preparation was 57 000 +/- 2 000 when determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The purified transferase catalysed the glucuronidation of approx. 0.6 mumol 4-nitrophenol min-1. mg-1 protein. Immunochemical analysis of this preparation further indicates that UDPglucuronosyltransferase has been purified to apparent homogeneity.
D000818:Animals; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D014453:Glucuronosyltransferase / Q000302:isolation & purification*; D005779:Immunodiffusion; D051379:Mice; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D051381:Rats
D003902:Detergents; D014453:Glucuronosyltransferase