Isolation and characterization of the plasma membrane of L-1210 cells. Iodination as a marker for the plasma membrane.
Mouse leukemia L-1210 cells were iodinated with 125I; this permitted the development of a method for the isolation of the plasma membranes. These show a 10- to 16-fold increase in the specific activity of 125I over that of the cell homogenate and a 20-fold increase in the specific activities of 5'-nucleotidase and alkaline phosphatase; 20-fold increase in the specific activities of 5'-nucleotidase and alkaline phosphatase; no mitochondrial or microsomal marker enzyme activities were detected. Sodium dodecyl sulfate gel electrophoresis of the plasma membranes shows approx. 40 peptides with molecular weights ranging from 10 000 to over 200 000; a polypeptide (Mr 50 000) predominates. Of 13 iodinated surface membrane proteins, the major radioactive peptide has a molecular weight of 85 000. The importance of the selection of the appropriate gel system for the analysis of membrane proteins is emphasized.
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005952:Glucose-6-Phosphatase / Q000032:analysis; D007455:Iodine; D007457:Iodine Radioisotopes; D007939:Leukemia L1210 / Q000032:analysis* / Q000201:enzymology; D051379:Mice; D008854:Microscopy, Electron; D009251:NADPH-Ferrihemoprotein Reductase / Q000032:analysis; D009708:Nucleotidases / Q000032:analysis; D013385:Succinate Dehydrogenase / Q000032:analysis
D007457:Iodine Radioisotopes; D003577:Cytochrome P-450 Enzyme System; D007455:Iodine; D013385:Succinate Dehydrogenase; D009251:NADPH-Ferrihemoprotein Reductase; D009708:Nucleotidases; D000469:Alkaline Phosphatase; D005952:Glucose-6-Phosphatase