Degradation of the sodium taurocholate cotransporting polypeptide (NTCP) by the ubiquitin-proteasome system.
The sodium taurocholate cotransporting polypeptide (Ntcp, Slc10a1) is the major uptake system for bile acids into liver cells. This study investigated the degradation of rat Ntcp and human NTCP by the ubiquitin-proteasome system (UPS). In stably transfected HepG2 cells, rat Ntcp was complex-glycosylated and localized at the plasma membrane. Inhibition of proteasomes by MG-132 or lactacystin led to the accumulation of intracellular Ntcp, a process dependent on de novo protein synthesis. Intracellular Ntcp was core-glycosylated, indicating an endoplasmic reticulum (ER) origin. Core-glycosylated Ntcp was found in cytosolic, detergent-insoluble deposits with characteristics of aggresomes: they co-localized with ubiquitin at the microtubule organization center and Ntcp from these deposits was polyubiquitinated. Transient transfections of Ntcp/NTCP induced intracellular deposits that co-localized with ubiquitin, even in the absence of proteasome inhibitors. Similarly, in livers of patients with progressive familial intrahepatic cholestasis, NTCP could be detected co-localized with ubiquitin in hepatocytes. We conclude that maturing Ntcp/NTCP is degraded by the ubiquitin-proteasome system at the level of ER-associated degradation (ERAD). An imbalance in the synthesis and degradation of NTCP at the level of the ER or alterations in the ERAD machinery might be the cause of intracellular NTCP deposits in transient transfections and in cholestatic livers.
Kühlkamp
Thomas
T
Department of Gastroenterology, Hepatology and Infectiology, Heinrich-Heine University, D-40225 Düsseldorf, Germany.
Keitel
Verena
V
Helmer
Angelika
A
Häussinger
Dieter
D
Kubitz
Ralf
R
D000818:Animals; D002462:Cell Membrane / Q000737:chemistry; D002478:Cells, Cultured; D002779:Cholestasis / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D003600:Cytosol / Q000737:chemistry; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology; D008099:Liver / Q000737:chemistry / Q000473:pathology; D026901:Membrane Transport Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D029363:Organic Anion Transporters, Sodium-Dependent; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D061988:Proteasome Inhibitors; D051381:Rats; D027981:Symporters; D025801:Ubiquitin / Q000032:analysis / Q000378:metabolism*
D015853:Cysteine Proteinase Inhibitors; D007976:Leupeptins; D026901:Membrane Transport Proteins; D029363:Organic Anion Transporters, Sodium-Dependent; D061988:Proteasome Inhibitors; D027981:Symporters; D025801:Ubiquitin; C071908:sodium-bile acid cotransporter; D046988:Proteasome Endopeptidase Complex; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde