Isolation of a rat liver plasma membrane fraction of probable canalicular origin. Preparative technique, enzymatic profile, composition, and solute transport.
A technique currently used for isolation of brush border membranes from renal and intestinal epithelium that involves vigorous tissue homogenization and sedimentation of non-luminal membranes in the presence of Mg2+ has been adapted to rat liver. Liver plasma membranes so prepared consisted almost exclusively of vesicles by electron microscopy, showed some contamination with endoplasmic reticulum and minimal contamination with mitochondria or Golgi by marker enzymes, were highly enriched in alkaline phosphatase, Mg2+-ATPase, and 5'-nucleotidase activity compared with homogenate, and showed little enrichment in (Na+, K+)-ATPase. Comparison of this enzymatic profile with cytochemical studies localizing (Na+, K+)-ATPase and alkaline phosphatase to the sinusoidal/lateral and canalicular membranes, respectively, suggested that these membranes were predominantly of canalicular origin. They had a lower (Na+ + K+)-ATPase specific activity, lower lipid content, and higher cholesterol to phospholipid molar ratio than a conventional plasma membrane preparation believed to be enriched in canaliculi. Moreover, it was possible to measure movement of D-[3H]glucose into an osmotically sensitive space bounded by these membrane vesicles.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000032:analysis* / Q000201:enzymology; D002784:Cholesterol / Q000032:analysis; D005947:Glucose / Q000378:metabolism; D008099:Liver / Q000032:analysis*; D008297:Male; D008854:Microscopy, Electron; D009994:Osmolar Concentration; D010743:Phospholipids / Q000032:analysis; D051381:Rats; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism
D010743:Phospholipids; D002784:Cholesterol; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D000254:Sodium-Potassium-Exchanging ATPase; D005947:Glucose