Application of high-performance liquid chromatographic techniques to the separation of ribosomal proteins of different organisms

• Published in: Journal of chromatography Vol. 317; p. 181
• Main Authors: Kamp, R M, Bosserhoff, A, Kamp, D, Wittmann-Liebold, B
• Format: Journal Article
• Language: English
• Published: Netherlands 01.01.1984
• Subjects: Amino Acid Sequence; Bacterial Proteins - analysis; Bacterial Proteins - isolation & purification; Chromatography, High Pressure Liquid - methods; Chromatography, Ion Exchange - methods; Electrophoresis, Polyacrylamide Gel - methods; Escherichia coli - metabolism; Euryarchaeota - analysis; Geobacillus stearothermophilus - analysis; Ribosomal Proteins - isolation & purification; Species Specificity
• DOI: 10.1016/S0021-9673(01)91658-9

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.