The subcellular localization of a bile acid glucosyltransferase in rat liver.

Formation of bile acid glucosides occurs in rat liver homogenate with a specific enzyme activity of 0.014 +/- 0.001 nmol per min per mg protein. Subcellular fractionation of rat liver by differential centrifugation revealed an enrichment of bile acid glucosyltransferase activity both in the mitochondrial-lysosomal fraction and in microsomes with a recovery of 38.8 +/- 4.6% and 37.7 +/- 1.7%, respectively, of enzyme activity in the homogenate. Subfractionation of the mitochondrial-lysosomal fraction after treatment of rats with Triton WR 1339 showed an almost exclusive association of bile acid glucosyltransferase activity with purified lysosomes ("tritosomes"). After subfractionation of microsomes by analytical gradients, bile acid glucosyltransferase was bimodally distributed with peaks at modal densities of 1.09 g/cm3 and 1.16 g/cm3, respectively. If microsomes were pretreated with pyrophosphate, a membrane perturbant known to strip ribosomes, only the peak of bile acid glucosyltransferase at higher density (1.16 g/cm3) and UDP-glucuronosyltransferase (marker of endoplasmic reticulum) shifted to a similar lower equilibrium density. Both enzymes were unaffected in their distribution by pretreatment of microsomes with digitonin. In contrast, markers of plasma membranes (5'-nucleotidase) and the Golgi-complex (galactosyltransferase) shifted to higher equilibrium densities after digitonin treatment, but were unaltered in their distribution after pyrophosphate. Bile acid glucosyltransferase activity in the lower density range with a peak at 1.09 g/cm3 did not show any association with the density distributions of known marker enzymes. In purified microsomal fractions obtained by preparative gradients, bile acid glucosyltransferase activity was enriched in enzyme activity by 1.4-fold in rough and by 2.3-fold in smooth endoplasmic reticulum, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)