Ribosomal proteins of E. coli and yeast were separated by gel filtration on dextran (Sephadex) and polyacrylamide (Bio-Gel) columns. Both gels revealed a valuable separation of the proteins. Finally only Bio-Gel columns were used, since their polyacrylamide matrix is more resistant to the applied organic acids.
The wide distribution of the molecular weights for both the E. coli and yeast ribosomal proteins was confirmed. E. coli ribosomal proteins were separated into three main groups by a single chromatography on Bio-Gel P-10. The same was true for yeast ribosomal proteins. Rechromatography of these protein groups resulted in a further valuable resolution. The fractionated proteins are recovered without any loss and they are very useful for further purification by other procedures, especially on a large scale basis.
KeywordsMolecular Weight Chromatography Purification Filtration Polyacrylamide
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- Delius, H., R. R. Traut, P. B. Moore, H. Noller, and P. Pearson: Studies on purified E. coli ribosomal proteins. In: Wissenschaftl. Konferenz der Ges. dtsch. Naturforscher und Ärzte, in Berlin 1967. International Symposium on “Molecular Genetics”. Berlin-Göttingen-Heidelberg: Springer in press.Google Scholar
- Duerre, J. A.: Fractionation of log phase Escherichia coli ribosomal proteins by gel filtration. Biochim. biophys. Acta (Amst.) 86, 490 (1964).Google Scholar
- Hamilton, M. G., and M. E. Ruth: Characterization of some of the proteins of the large subunit of rat liver ribosomes. Biochemistry 6, 2585 (1967).Google Scholar
- Kaltschmidt, E., V. Rudloff, G. Stöffler, A. Chersi, M. Dzionara, D. Donner, and H. G. Wittmann: Ribosomal proteins of E. coli and yeast. In: Wissenschaftl. Konferenz der Ges. dtsch. Naturforscher und Ärzte, in Berlin 1967. International Symposium on “Molecular Genetics”. Berlin-Göttingen-Heidelberg: Springer in press.Google Scholar
- Lowry, O. H., N. J. Rosebrough, A. L. Farr, and R. J. Randall: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265 (1951).Google Scholar
- Möller, W., and A. Chrambach: Physical heterogeneity of the ribosomal proteins from Escherichia coli. J. molec. Biol. 23, 377 (1967).Google Scholar
- Moore, S., and W. H. Stein: Photometric ninhydrin method for use in the chromatography of amino acids. J. biol. Chem. 176, 367 (1948).Google Scholar
- Morris, D. L.: Quantitative determination of carbohydrates with Dreywood's anthrone reagent. Science 107, 254 (1948).Google Scholar
- Reisfeld, R. A., U. J. Lewis, and D. E. Williams: Disc electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature (Lond.) 195, 281 (1962).Google Scholar
- Shapiro, A. L., E. Vinuela, and J. V. Maizel jr.: Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem. biophys. Res. Commun. 28, 815 (1967).Google Scholar
- Traut, R. R.: Acrylamide gel electrophoresis of radioactive ribosomal protein. J. molec. Biol. 21, 571 (1966).Google Scholar
- Waller, J. P., and J. I. Harris: Studies on the composition of the protein from Escherichia coli ribosomes. Proc: nat. Acad. Sci. (Wash.) 47, 18 (1961).Google Scholar