Isolation of the plasma membrane from corneal endothelial cells.
The plasma membtanes of normal rabbit endothelail cells were isolated by the use of an aqueous two-phase polymer system. The membrane fraction was identified by electron microscopy, and a fivefold to eightfold increase in the specific activity of two plasma membrane markers. Na+-K+-ATPase and 5'nucleotidase was found. Recovery of the enzyme markers averaged 45% and 22%, respectively. Analysis of the purified membranes for glucose-6-phosphatase, a marker for endoplasmic reticulum, showed no contamination by this structure. This method for cell membrane characterization is promising in determining the enzymatic alterations of diseased corneas.
D000251:Adenosine Triphosphatases / Q000032:analysis; D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure; D003315:Cornea / Q000201:enzymology / Q000648:ultrastructure*; D005952:Glucose-6-Phosphatase / Q000032:analysis; D008274:Magnesium / Q000032:analysis; D008854:Microscopy, Electron; D009708:Nucleotidases / Q000032:analysis; D011817:Rabbits; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis
D009708:Nucleotidases; D005952:Glucose-6-Phosphatase; D000251:Adenosine Triphosphatases; D000254:Sodium-Potassium-Exchanging ATPase; D008274:Magnesium