Kinetic analysis of UDP-glucuronosyltransferase in bilirubin conjugation by encapsulated hepatocytes for transplantation into Gunn rats.
Kinetic analysis of the enzyme UDP-glucuronosyltransferase (UDPGT), responsible for the conjugation of bilirubin, suggests that it is a multisubunit enzyme in which there is cooperative binding of the substrate to the subunits. The binding of bilirubin to UDPGT shows positive cooperativity with an apparent Hill coefficient of 2.9. The binding of UDP-glucuronic acid (UDPGA) exhibits kinetics with mixed cooperativity with an apparent Hill coefficient of 4.028. Homogenized rat hepatocytes, intact hepatocytes, and hepatocytes encapsulated in alginate-polylysine-alginate artificial cells, when incubated with bilirubin (1.6 mM) and UDPGA (20 mM), can form monoconjugated and diconjugated bilirubin. However, the presence of the artificial membrane offers some mass transfer resistance. The intraperitoneal transplantation into the Gunn rat of free and microencapsulated Wistar rat hepatocytes shows that both are equally effective in lowering the serum bilirubin. Thus, the membrane did not contribute to a lowering of efficacy after transplantation.
Bruni
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Artificial Cells and Organs Research Centre, Faculty of Medicine, McGill University, Montreal, Quebec, Canada.
Chang
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TM
D000464:Alginates; D000818:Animals; D001663:Bilirubin / Q000032:analysis / Q000097:blood / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D014453:Glucuronosyltransferase / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000166:cytology* / Q000201:enzymology; D016031:Liver Transplantation / Q000295:instrumentation / Q000379:methods*; D008297:Male; D008567:Membranes, Artificial; D008863:Microspheres; D010537:Peritoneum; D011107:Polylysine; D051381:Rats; D011919:Rats, Inbred Strains; D017208:Rats, Wistar; D014184:Transplantation, Homologous
D000464:Alginates; D008567:Membranes, Artificial; D011107:Polylysine; D014453:Glucuronosyltransferase; D001663:Bilirubin