Comparison of the effect of various methods of dissociation on the protein composition of rat liver ribosomal subunits.
Rat liver ribosomes were dissociated into subunits using EDTA, sodium pyrophosphate, high concentrations of KC1, as well as by incubation with puromycin in presence of 0.5 M KC1. The subunits obtained were analyzed using the density gradient centrifugation technique and their ribosomal proteins were separated by means of two-dimensional polyacrylamide gel electrophoresis. The ribosomal protein patterns of the two subunits isolated using each of the dissociating method were compared to the protein patterns of monosomes prepared by puromycin treatment alone. Our results revealed that the use of chelating agents to dissociate the ribosomes resulted in the loss of some ribosomal proteins from the small subunit. On the other hand, the use of KC1 in high concentrations to dissociate the ribosomes did not appear to cause any major loss of proteins from the ribosomes except for some acidic proteins.
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D011756:Diphosphates; D004492:Edetic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000032:analysis* / Q000648:ultrastructure; D008297:Male; D011189:Potassium Chloride; D011691:Puromycin; D051381:Rats; D012269:Ribosomal Proteins / Q000032:analysis*; D012270:Ribosomes / Q000032:analysis*
D011756:Diphosphates; D012269:Ribosomal Proteins; D011691:Puromycin; D011189:Potassium Chloride; D004492:Edetic Acid