Liver alcohol dehydrogenase is degraded by the ubiquitin-proteasome pathway.
Dihydrotestosterone (DHT) decreases rat liver alcohol dehydrogenase (ADH) due principally to an increased rate of degradation of the enzyme. The pathway of degradation of ADH was investigated. Exposure of hepatocytes in culture to lactacystin or to MG132, which are inhibitors of the ubiquitin-proteasome pathway of protein degradation, resulted in higher ADH. Furthermore, both lactacystin and MG132 prevented the decrease in ADH caused by DHT. By contrast, the lysosomal proteolytic inhibitors 3-methyladenine and leupeptin as well as inhibitors of the calcium-activated neutral protease calpain system had no effect on ADH in the absence or presence of DHT. ADH isolated by immunoprecipitation from hepatocytes exposed to DHT reacted specifically with anti-ubiquitin antibody. Ubiquitinated ADH was also demonstrated in hepatocytes exposed to MG132. The combination of DHT and MG132 resulted in more ubiquitinated ADH than exposure to either compound alone. These results suggest that the ubiquitin-proteasome pathway plays a role in the degradation of ADH and in the enhanced degradation of this enzyme by DHT.
Mezey
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Department of Medicine, Johns Hopkins University School of Medicine, 921 Ross Research Building, 720 Rutland Avenue, Baltimore, MD 21205, USA. emezey@jhmi.edu
Rennie-Tankersley
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Potter
J J
JJ
D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000426:Alcohol Dehydrogenase / Q000378:metabolism*; D000818:Animals; D002154:Calpain / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D013196:Dihydrotestosterone / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D007976:Leupeptins / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D009097:Multienzyme Complexes / Q000378:metabolism*; D011233:Precipitin Tests; D046988:Proteasome Endopeptidase Complex; D051381:Rats; D017207:Rats, Sprague-Dawley; D014452:Ubiquitins / Q000378:metabolism*
D015853:Cysteine Proteinase Inhibitors; D007976:Leupeptins; D009097:Multienzyme Complexes; D014452:Ubiquitins; D013196:Dihydrotestosterone; C067713:lactacystin; C025946:3-methyladenine; D000426:Alcohol Dehydrogenase; D002154:Calpain; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; C032854:leupeptin; D000225:Adenine; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde; D000111:Acetylcysteine