Association of canalicular membrane enzymes with bile acid micelles and lipid aggregates in human and rat bile.
This study was undertaken to gain insights into the characteristics of the polymolecular association between canalicular membrane enzymes, bile acids, cholesterol and phospholipids in bile and into the celular mechanisms whereby the enzymes are secreted into bile. With this purpose, we studied the distribution of bile acids, cholesterol, phospholipids, proteins and representative canalicular membrane enzymes (alkaline phosphatase, 5'-nucleotidase and gamma-glutamyl transpeptidase), which can be considered specific marker constituents, in bile fractions enriched in phospholipid-cholesterol lamellar structures (multilamellar and unilamellar vesicles) and bile acid-mixed micelles. These fractions were isolated by ultracentrifugation from human hepatic bile, normal rat bile and bile of rats treated with diosgenin, a steroid that induces a marked increase in biliary cholesterol secretion, and were characterized by density, lipid composition and transmission electron microscopy. These studies demonstrate that alkaline phosphatase, 5'-nucleotidase and gamma-glutamyl transpeptidase are secreted into both human and rat bile where they are preferentially associated with bile acid-mixed micelles, suggesting a role for bile acids in both release of these enzymes and lipids from the canalicular membrane and solubilization in bile. In addition, heterogeneous association of these enzymes with nonmicellar, lamellar structures in human and rat bile is consistent with the hypothesis that processes independent of the detergent effects of bile acids might also result in the release of specific intrinsic membrane proteins into bile.
Accatino
L
L
Departamento de Gastroenterología, Facultad de Medicina, P. Universidad Católica de Chile, Santiago.
Pizarro
M
M
Solís
N
N
Koenig
C S
CS
D015720:5'-Nucleotidase / Q000032:analysis; D000368:Aged; D000369:Aged, 80 and over; D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D001646:Bile / Q000737:chemistry* / Q000201:enzymology*; D001647:Bile Acids and Salts; D001648:Bile Canaliculi / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D006801:Humans; D008055:Lipids; D008297:Male; D008566:Membranes / Q000201:enzymology / Q000648:ultrastructure; D008823:Micelles; D008875:Middle Aged; D051381:Rats; D017208:Rats, Wistar; D013045:Species Specificity; D005723:gamma-Glutamyltransferase / Q000032:analysis
D001647:Bile Acids and Salts; D008055:Lipids; D008823:Micelles; D005723:gamma-Glutamyltransferase; D000469:Alkaline Phosphatase; D015720:5'-Nucleotidase