A rapid and preparative method for the separation of yeast ribosomal proteins by using high-performance liquid chromatography

Abstract

Ribosomal proteins from the yeast Saccharomyces cerevisiae were separated, on a preparative scale, by ion-exchange h.p.l.c. Proteins from the small and large ribosomal subunits were resolved, respectively, into 33 and 23 peaks, and most of the proteins present in these peaks were identified by using one- and two-dimensional gel electrophoresis. Several of the peaks appeared to contain a single protein uncontaminated by other species. Ribosomal proteins were also separated by using reverse-phase h.p.l.c. Analysis of the peaks resolved indicated that the order of elution for the proteins of both ribosomal subunits is, in certain cases, different for each of the two h.p.l.c. techniques used. Thus a combination of the two chromatographic methods employed here has the potential to facilitate the rapid and preparative separation of each of the proteins present in yeast ribosomes.

tRNA-Assisted Overproduction of Eukaryotic Ribosomal Proteins

“…This may be due to difficulties in the preparative disassembly and isolation of individual ribosomal components or in the production of recombinant eukaryotic r-proteins (17,18). In keeping with the latter possibility, we noticed that r-proteins from various eukaryotes are expressed at very low levels in E. coli.…”

Synthesis of ribosomes in Saccharomyces cerevisiae.

“…There are approximately 75 proteins in a yeast cytoplasmic ribosome. These have been studied mostly by twodimensional polyacrylamide gel techniques (11,100,118,187), although high-performance liquid chromatography is currently the method of choice for the purification of individual proteins (165). Unfortunately, both for the reader and for the workers in the field, there are three systems of nomenclature for the ribosomal proteins, which have been only partly correlated (119,131,183).…”