ATP-dependent bile-salt transport in canalicular rat liver plasma-membrane vesicles.
The present study identifies and characterizes a novel ATP-dependent bile-salt transport system in isolated canalicular rat liver plasma-membrane (cLPM) vesicles. ATP (1-5 mM) stimulated taurocholate uptake into cLPM vesicles between 6- and 8-fold above equilibrium uptake values (overshoot) and above values for incubations in the absence of ATP. The ATP-dependent portion of taurocholate uptake was 2-fold higher in the presence of equilibrated KNO3 as compared with potassium gluconate, indicating that the stimulatory effect of ATP was not due to the generation of an intravesicular positive membrane potential. Saturation kinetics revealed a very high affinity (Km approximately 2.1 microM) of the system for taurocholate. The system could only minimally be stimulated by nucleotides other than ATP. Furthermore, it was preferentially inhibited by conjugated univalent bile salts. Further strong inhibitory effects were observed with valinomycin, oligomycin, 4,4'-di-isothiocyano-2,2'-stilbene disulphonate, sulphobromophthalein, leukotriene C4 and N-ethylmaleimide, whereas nigericin, vanadate, GSH, GSSG and daunomycin exerted only weak inhibitory effects or none at all. These results indicate the presence of a high-affinity primary ATP-dependent bile-salt transport system in cLPM vesicles. This transport system might be regulated in vivo by the number of carriers present at the perspective transport site(s), which, in addition to the canalicular membrane, might also include pericanalicular membrane vesicles.
Stieger
B
B
Department of Medicine, University Hospital, Zurich, Switzerland.
O'Neill
B
B
Meier
P J
PJ
D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism / Q000493:pharmacokinetics*; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism / Q000502:physiology; D006868:Hydrolysis; D007700:Kinetics; D008099:Liver / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D051381:Rats; D011919:Rats, Inbred Strains; D013379:Substrate Specificity; D013656:Taurocholic Acid / Q000378:metabolism / Q000493:pharmacokinetics; D014316:Tritium
D001647:Bile Acids and Salts; D014316:Tritium; D013656:Taurocholic Acid; D000255:Adenosine Triphosphate