Isolation of yeast ribosomal proteins L3 and L2 for immunological studies.
We report on a rapid method for the isolation and purification of the yeast ribosomal proteins L3 and L2 using a simple instrumentation. Preparative dodecyl sulfate polyacrylamide gel electrophoresis was applied to the separation of cytoplasmatic ribosomal proteins of the large subunit from the yeast Saccharomyces cerevisiae. The polypeptides were removed from gel slices by electrophoretic elution. Subsequent analytical electrophoresis showed groups of proteins in all but two fractions. The latter were further analysed by a two-dimensional gel electrophoresis system which disclosed the purity of two polypeptides. They were identified as L3 and L2. Their molecular masses were 51.5 and 44 kDa as estimated from the gels. A possible application to the isolation of other yeast ribosomal proteins is discussed. An antiserum against the polypeptide L3 was raised in a rabbit. Applying an enzyme-linked immunosorbent assay (ELISA) we were able to determine the relative antibody concentration. Its specificity was demonstrated by immunoblotting.
D000908:Antibodies, Fungal / Q000276:immunology; D005656:Fungal Proteins / Q000276:immunology / Q000302:isolation & purification*; D000090822:Ribosomal Protein L3; D012269:Ribosomal Proteins / Q000276:immunology / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000032:analysis* / Q000276:immunology
D000908:Antibodies, Fungal; D005656:Fungal Proteins; D000090822:Ribosomal Protein L3; D012269:Ribosomal Proteins; C029466:ribosomal protein L2