Localization of the ecto-ATPase (ecto-nucleotidase) in the rat hepatocyte plasma membrane. Implications for the functions of the ecto-ATPase.
The surface distribution of the plasma membrane Ca2+ (Mg2+)-ATPase (ecto-ATPase) in rat hepatocytes was determined by several methods. 1) Two polyclonal antibodies specific for the ecto-ATPase were used to examine the distribution of the enzyme in frozen sections of rat liver by immunofluorescence. Fluorescent staining was observed at the bile canalicular region of hepatocytes. 2) Plasma membranes were isolated from the canalicular and sinusoidal regions of rat liver. The specific activity of ecto-ATPase in the canalicular membranes was 22 times higher than that of sinusoidal membranes. The enrichment of the ecto-ATPase activity in the canalicular membrane is closely parallel to that of two other canalicular membrane markers, gamma-glutamyltranspeptidase and leucine aminopeptidase. 3) By immunoblots with polyclonal antibodies against the ecto-ATPase and the Na+,K+-ATPase, it was found that the ecto-ATPase protein was only detected in canalicular membranes and not in sinusoidal membranes, while the Na+,K+-ATPase protein was only detected in sinusoidal membranes and not in canalicular membranes. These results indicate that the ecto-ATPase is enriched in the canalicular membranes of rat hepatocytes.
Lin
S H
SH
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, Massachusetts 02138.
D000818:Animals; D000252:Calcium-Transporting ATPases / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D002458:Cell Fractionation; D002462:Cell Membrane / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D007106:Immune Sera; D015151:Immunoblotting; D008099:Liver / Q000201:enzymology*; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D011817:Rabbits; D051381:Rats
D007106:Immune Sera; D008565:Membrane Proteins; D000252:Calcium-Transporting ATPases