Isolation and partial characterization of the major glycoprotein from the plasma membranes of AH-66 hepatoma cells.
A major glycoprotein of the plasma membranes of AH-66 hepatoma ascites cells was isolated in essentially pure form and in milligram amounts. The plasma membranes were solubilized with a solution containing both 0.3 M lithium diiodosalycylate and 0.2% cetylpyridinium chloride, and further extracted with 50% phenol, followed by gel filtration on Sepharose 6B in the presence of 0.1% Ammonyx-LO at pH 8.0. The apparent molecular weight of the purified glycoprotein was estimated to be 165 000 in 5.6% polyacrylamide gels, of which 54% was carbohydrate and 46% was protein. The chemical composition of the glycoprotein resembles glycophorin A from human erythrocyte membranes in that it has a high content of N-acetylgalactosamine, N-acetylglucosamine, galactose and sialic acid and a particularly large proportion of serine, threonine, aspartic acid and glutamic acid.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000302:isolation & purification*; D008114:Liver Neoplasms, Experimental / Q000032:analysis*; D008565:Membrane Proteins / Q000302:isolation & purification*; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000302:isolation & purification*; D051381:Rats
D000596:Amino Acids; D006023:Glycoproteins; D008565:Membrane Proteins; D009363:Neoplasm Proteins