The location of proteins labeled by the 125I-lactoperoxidase system in the NIL 8 hamster fibroblast.
NIL 8 hamster fibroblast cells were labeled by lactoperoxidase-catalyzed iodination. Their membranes were fractionated by sedimentation-rate and isopycnic zonal centrifugation. All the iodinated proteins except the very prominently labeled high molecular weight protein (greater than 200,000 daltons) were located in a fraction identified enzymically and compositionally as plasma membrane. The high molecular weight protein that was previously shown to be sensitive to virus transformation (Hynes, 1973) is concentrated in a very high density particle (rho equals 1.253-1.259) which contains mainly carbohydrate and protein and very low levels of lipid. 5'-nucleotidase was the only enzyme reproducibly demonstrated in this fraction, and electron micrographs revealed a predominantly amorphous morphology together with a few membraneous structures. The iodine label in this fraction was very sensitive to trypsinization prior to homogenization. All the available evidence indicates that this fraction is derived from the surface coat. Mitochondria, nuclei, and soluble protein were labeled to an insignificant extent. The presence of the iodinated surface proteins associated with the endoplasmic reticulum fraction is discussed in the light of these results.
D000135:Acid Phosphatase / Q000032:analysis; D000251:Adenosine Triphosphatases / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002374:Catalase / Q000032:analysis; D002458:Cell Fractionation; D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis; D002467:Cell Nucleus / Q000032:analysis; D002499:Centrifugation, Density Gradient; D006224:Cricetinae; D003579:Cytochrome Reductases / Q000032:analysis; D008058:Dihydrolipoamide Dehydrogenase / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000032:analysis; D005347:Fibroblasts / Q000032:analysis*; D007457:Iodine Radioisotopes; D007931:Leucyl Aminopeptidase / Q000032:analysis; D008055:Lipids / Q000032:analysis; D008854:Microscopy, Electron; D008928:Mitochondria / Q000032:analysis; D009699:N-Glycosyl Hydrolases / Q000032:analysis; D009696:Nucleic Acids / Q000032:analysis; D010544:Peroxidases; D011506:Proteins / Q000302:isolation & purification*; D013385:Succinate Dehydrogenase / Q000032:analysis
D002241:Carbohydrates; D007457:Iodine Radioisotopes; D008055:Lipids; D009696:Nucleic Acids; D011506:Proteins; D010544:Peroxidases; D002374:Catalase; D013385:Succinate Dehydrogenase; D003579:Cytochrome Reductases; D008058:Dihydrolipoamide Dehydrogenase; D000135:Acid Phosphatase; D009699:N-Glycosyl Hydrolases; D007931:Leucyl Aminopeptidase; D000251:Adenosine Triphosphatases