Effect of albumin on the in vitro conjugation of bilirubin by rat liver microsomes.
These studies were carried out to determine whether bovine serum albumin (BSA), which is usually included in the incubation mixture for the in vitro determination of bilirubin-UDP-glucuronyl transferase (GT) activity, affects GT activity. Using bilirubin as substrate, addition of BSA to the enzyme reaction mixture at concentrations varying from 2 to 30 mg/ml resulted in a dose-related inhibition of "native" GT activity of rat liver microsomes. When detergent-activated enzyme was employed, increasing concentrations of BSA also required higher concentrations of deoxycholate, digitonin, or Triton X-100 to produce maximal bilirubin conjugation. Low BSA concentrations (2 mg/ml) prevented enzyme activation by both detergents and UDP-N-acetyl glucosamine. When BSA was omitted and bilirubin dissolved in dimethyl sulfoxide, UDP-N-acetyl glucosamine failed to enhance GT activity, and activation by detergents was only 15-25% of that observed in the presence of optimal concentrations of BSA. When rat albumin was substituted for BSA, a similar dose-related inhibition of in vitro bilirubin conjugation by untreated microsomes was observed, although at any given albumin concentration, GT activity was lower with rat than with bovine albumin. Additionally, both detergents and UDP-N-acetyl glucosamine produced similar GT activation regardless of the rat albumin concentration. Finally, these effects of BSA and rat albumin could not be reproduced when beta-lactoglobulin was employed and/or when p-nitrophenol was the acceptor substrate of GT. These findings indicate that albumin, in particular BSA, profoundly and selectively influences the in vitro activity of microsomal GT toward bilirubin as the acceptor substrate.
D000818:Animals; D001663:Bilirubin / Q000378:metabolism*; D003840:Deoxycholic Acid / Q000494:pharmacology; D004072:Digitonin / Q000494:pharmacology; D004789:Enzyme Activation; D014453:Glucuronosyltransferase; D006602:Hexosyltransferases / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D012710:Serum Albumin, Bovine / Q000494:pharmacology*; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000494:pharmacology
C019787:bilirubin glucuronoside glucuronosyltransferase; D003840:Deoxycholic Acid; D012710:Serum Albumin, Bovine; D014537:Uridine Diphosphate N-Acetylglucosamine; D006602:Hexosyltransferases; D014453:Glucuronosyltransferase; D004072:Digitonin; D001663:Bilirubin