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,...

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Bibliographic Details
Published in:Journal of Chromatography A 1984-01, Vol.317, p.181-192
Main Authors: Kamp, Roza Maria, Bosserhoff, Armin, Kamp, Detlev, Wittmann-Liebold, Brigitte
Format: Article
Language:English
Subjects:
Bacillus stearothermophilus
Escherichia coli
high-performance liquid chromatography
Methanococcus vannielii
ribosomal proteins
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Summary: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 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.
ISSN:0021-9673
DOI:10.1016/S0021-9673(01)91658-9