ENHANCEMENT OF MDR2-MEDIATED PHOSPHATIDYLCHOLINE TRANSLOCATION BY THE BILE-SALT TAUROCHOLATE - IMPLICATIONS FOR HEPATIC BILE FORMATION

Expression of the Mdra-protein in secretory vesicules (SVs) from the yeast mutant sec6-4 causes a time- and temperature-dependent enhancement of phosphatidylcholine (PC) translocation from the outer to the inner leaflet of the SV lipid bilayer. We show that this activity is independent of changes either in the membrane potential or the pH gradient (inside positive) generated in these SVs by the yeast proton-translocating PMA1 ATPase. However, loading of the SVs with the primary bile salt taurocholate results in an apparent enhancement of Mdr2-mediated PC translocation activity. Reducing the intravesicular taurocholate (TC) concentration by dissipating the electrochemical potential across the SV membranes eliminates the enhancing effect of TC. Three Lines of evidence suggest that the enhanced Mdr2-mediated PC translocation activity is not caused by a regulatory effect of TC on Mdr2 but rather reflected the formation of TC/PC aggregates or micelles in the lumen of SVs. First, significantly higher detergent concentrations are required to reveal the fluorescence of (7-niaro-2-1,3-benzoxadiazol-4-yl)amino- molecules translocated in Mdr2-SV under conditions of TC stimulation than under control conditions; second, the nonmicelle-forming bile salt taurodehydrocholate does not cause enhancement of PC translocation in Mdr2-SVs; third, enzyme marker studies indicate that TC behaves as a potent lipid solubilizer directly extracting PC molecules out of the bilayer without causing leakage. This results in the formation of intravesicular aggregates or mixed micelles, and provokes the apparent stimulation of Mdr2 activity. These data demonstrate a unique relationship between Mdr2, PC, and TC in the process of bile formation and secretion.