Isolation of Yeast Ribosomal Proteins L3 and L2 for Immunological Studies

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Abstract
Preparative dodecyl sulfate polyacrylamide gel electrophoresis was applied to the separation of cytoplasmatic ribosomal proteins of the large subunit from the yeast S. cerevisiae. The polypeptides were removed from gel slices by electrophoretic elution. Subsequent analytical electrophoresis showed groups of proteins in all but 2 fractions. The latter were further analyzed by a 2-dimensional gel electrophoresis system which disclosed the purity of 2 polypeptides. They were identified as L3 and L2. Their molecular masses were 51.5 and 44 kDa [kilodalton] 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. With enzyme-linked immunosorbent assay (ELISA) the relative antibody concentration was determined. Its specifity was demonstrated by immunoblotting.