Application of high-performance liquid chromatographic techniques to the separation of ribosomal proteins of different organisms.
The ribosomal proteins from Escherichia coli, Bacillus stearothermophilus and Methanococcus vannielii were separated by size-exclusion, ion-exchange and reversed-phase high-performance liquid chromatography (HPLC), employing new column materials, different gradient systems, and preparative columns, respectively. The purity of the isolated proteins was analysed by one- and two-dimensional gel electrophoresis and by direct micro-sequencing. The separation of ribosomal proteins could be improved by employing propanol gradients in combination with Vydac reversed-phase columns. From the E. coli ribosome, fifteen S and twenty-three L proteins were isolated in sequencer purity by this method. In addition, ion-exchange HPLC was proven to be useful for isolating ribosomal proteins under native conditions: six S proteins and sixteen L proteins from E. coli could be purified. Some of these proteins were not isolated by the reversed-phase procedures, e.g. proteins L9, L14 and L21.
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D019605:Euryarchaeota / Q000032:analysis; D001411:Geobacillus stearothermophilus / Q000032:analysis; D012269:Ribosomal Proteins / Q000302:isolation & purification*; D013045:Species Specificity
D001426:Bacterial Proteins; D012269:Ribosomal Proteins