Accelerated degradation of mislocalized UDP-glucuronosyltransferase family 1 (UGT1) proteins in Gunn rat hepatocytes.
Gunn rat is a hyperbilirubinemic rat strain that is inherently deficient in the activity of UDP-glucuronosyltransferase form 1A1 (UGT1A1). A premature termination codon is predicted to produce truncated UGT1 proteins that lack the COOH-terminal 116 amino acids in Gunn rat. Pulse-chase experiments using primary cell cultures showed that the truncated UGT1A1 protein in Gunn rat hepatocytes was synthesized similarly to wild-type UGT1A1 protein in normal Wistar rat hepatocytes. However, the truncated UGT1A1 protein was degraded rapidly with a half-life of about 50 min, whereas the wild-type UGT1A1 protein had a much longer half-life of about 10 h. The rapid degradation of truncated UGT1A1 protein was inhibited partially but not completely by treating Gunn rat hepatocytes with proteasome inhibitors such as carbobenzoxy-Leu-Leu-leucinal and lactacystin. By contrast, neither the lysosomal cysteine protease inhibitor nor the calpain inhibitor slowed the degradation. Our findings show that the absence of UGT1 protein from Gunn rat hepatocytes is due to rapid degradation of the truncated UGT1 protein by the proteasome and elucidate the molecular basis underlying the deficiency in bilirubin glucuronidation.
Emi
Yoshikazu
Y
Department of Life Science, Faculty of Science, Himeji Institute of Technology, Harima Science Park City, Hyogo 678-1297, Japan. emys@sci.himeji-tech.ac.jp
Omura
Satoshi
S
Ikushiro
Shin-ichi
S
Iyanagi
Takashi
T
D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors; D014453:Glucuronosyltransferase / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D006023:Glycoproteins / Q000494:pharmacology; D006207:Half-Life; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D007976:Leupeptins / Q000494:pharmacology; D008297:Male; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism; D009154:Mutation; D009842:Oligopeptides / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011911:Rats, Gunn; D017208:Rats, Wistar; D013347:Subcellular Fractions; D000099237:UDP-Galactose Translocators; D000099148:UGT1A1 Enzyme
D000111:Acetylcysteine; D003546:Cysteine Endopeptidases; D015853:Cysteine Proteinase Inhibitors; D014453:Glucuronosyltransferase; D006023:Glycoproteins; D007976:Leupeptins; D009004:Monosaccharide Transport Proteins; D009097:Multienzyme Complexes; D009842:Oligopeptides; D046988:Proteasome Endopeptidase Complex; D012333:RNA, Messenger; D000099237:UDP-Galactose Translocators; D000099148:UGT1A1 Enzyme; C499946:Ugt1a1 protein, rat; C039304:calpain inhibitors; C069861:calpain inhibitor 2; C067713:lactacystin; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde