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Characterisation of eukaryotic ribosomal proteins

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Summary

A simple method of two-dimensional polyacrylamide gel electrophoresis is described which affords: (1) high resolution of eukaryotic ribosomal proteins; (2) good recovery of protein in the transfer from first to second dimension; and (3) characterisation of the separated proteins in terms of molecular weights and other electrophoretic properties. Using this method, we have characterised 70 proteins in rabbit reticulocyte ribosomes, 30 from the small subunit and 40 from the large subunit. The molecular weight distribution is compared with those obtained by other authors after fractionation of the proteins in two dimensions.

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References

  • Bickle, T. A., Traut, R. R.: Differences in size and number of 80 S and 70 S ribosomal proteins by dodecyl sulfate gel electrophoresis. J. biol. Chem. 246, 6828–6834 (1971)

    Google Scholar 

  • Blobel, G.: Isolation of a 5S ribosomal RNA-protein complex from mammlian ribosomes. Proc. nat. Acad. Sci. (Wash.) 68, 1881–1885 (1971)

    Google Scholar 

  • Chaterjee, S. K., Kazemie, M., Matthaei, H.: Studies on rabbit reticulocyte ribosomes. II. Separation of the ribosomal proteins by two-dimensional electrophoresis. Hoppe-Seylers Z. physiol. Chem. 354, 481–486 (1973)

    Google Scholar 

  • Dzionara, M., Kaltschmidt, E., Wittmann, H.-G.: Ribosomal proteins. XIII. Molecular weights of isolated ribosomal proteins of Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 67, 1909–1913 (1970)

    Google Scholar 

  • Gressner, A. M., Wool, I. G.: The phosphorylation of liver ribosomal proteins in vivo: Evidence that only a single small subunit protein (S6) is phosphorylated. J. biol. Chem. 249, 6917–6925 (1974)

    Google Scholar 

  • Hardy, S. J. S., Kurland, C. G., Voynow, P., Mora, G.: The ribosomal proteins of Escherichia coli. I. Purification of the 30S ribosomal proteins. Biochem. 8 2897–2905 (1969)

    Google Scholar 

  • Hindennach, I., Kaltschmidt, E., Wittmann, H.-G.: Ribosomal proteins. Isolation of proteins from 50S ribosomal subunits of Escherichia coli. Europ. J. Biochem. 23, 12–16 (1971b)

    Google Scholar 

  • Hindennach, I., Stöffler, G., Wittmann, H.-G.: Ribosomal proteins. Isolation of the proteins from 30S ribosomal subunits of Escherichia coli. Europ. J. Biochem. 23, 7–11 (1971a)

    Google Scholar 

  • Howard, G. A., Traut, R. R.: A modified two-dimensional gel system for the separation and radioautography of microgram amounts of ribosomal proteins. Methods in Enzymol. 30, 526–539 (1974)

    Google Scholar 

  • Howard, G. A., Traugh, J. A., Croser, E. A., Traut, R. R.: Ribosomal proteins from rabbit reticulocytes: number and molecular weights of proteins from ribosomal subunits. J. molec. Biol. 93, 391–404 (1975)

    Google Scholar 

  • Hultin, T., Sjöqvist, A.: Analogies in protein patterns and among ribosomes from different classes of vertebrates. Comp. Biochem. Physiol. 40B, 1011–1027 (1971)

    Google Scholar 

  • Huynh-Van-Tan, Delauny, J., Schapira, G.: Eukaryotic ribosomal proteins. Two-dimensional electrophoretic studies. FEBS Letters 17, 163–167 (1971)

    Google Scholar 

  • Kaltschmidt, E., Wittmann, H. G.: Ribosomal proteins. VII. Two-dimensional polyacrylamide gel electrophoresis for finger-printing of ribosomal proteins. Analyt. Biochem. 30, 401–412 (1970a)

    Google Scholar 

  • Kaltschmidt, E., Wittmann, H.-G.: Ribosomal proteins. XII. Number of proteins in small and large ribosomal subunits of Escherichia coli as determined by two-dimensional gel electrophoresis. Proc. nat. Acad. Sci. (Wash.) 67, 1276–1282 (1970b)

    Google Scholar 

  • King, H. W. S., Gould, H. J., Shearman, J. J.: Molecular weight distribution of proteins in rabbit reticulocyte ribosomal subunits. J. molec. Biol. 61, 143–156 (1971)

    Google Scholar 

  • Leboy, P. S., Cox, E. C., Flaks, J. G.: The chromosomal site specifying a ribosomal protein in Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 52, 1367–1374 (1964)

    Google Scholar 

  • Lin, A., Wool, I. G.: The molecular weights of rat liver ribosomal proteins determined by “three-dimensional” polyacrylamide gel electrophoresis. Molec. gen. Genet. 134, 1–6 (1974)

    Google Scholar 

  • Loening, U. E.: The fractionation of high molecular weight RNA by polyacrylamide gel electrophoresis. J. Biochem. 102, 251–257 (1967)

    Google Scholar 

  • Martin, T. E., Wool, I. G.: Formation of active hybrids from subunits of muscle ribosomes from normal and diabetic rats. Proc. nat. Acad. Sci. (Wash.) 60, 569–574 (1968)

    Google Scholar 

  • Martini, O. H. W.: Eukaryotic ribosomal proteins. Ph.D. Thesis, London University (1974)

  • Martini, O. H. W., Gould, H. J.: Enumeration of rabbit reticulocyte ribosomal proteins. J. molec. Biol. 62, 403–405 (1971)

    Google Scholar 

  • Martini, O. H. W., Gould, H. J.: Phosphorylation of rabbit reticulocyte ribosomal proteins in vitro. Biochim. biophys. Acta (Amst.) 295, 621–629 (1973)

    Google Scholar 

  • Martini, O. H. W., Gould, H. J.: Molecular weight distribution of ribosomal proteins from different vertebrate species. Molec. gen. Genet. 142, 317–331 (1975)

    Google Scholar 

  • Martini, O. H. W., Gould, H. J., King, H. W. S.: 80S ribosomal proteins, in The structure and function of eukaryotic ribosomes. Biochem. Soc. Symp. 37, 51–68 (1973)

    Google Scholar 

  • Martini, O. H. W., Temkin, R., Jones, A., Riley, K., Gould, H. J.: Quantitation of eukaryotic ribosomal proteins separated by two-dimensional polyacrylamide gel electrophoresis. FEBS Letters 56, 205–211 (1975)

    Google Scholar 

  • Mets, L. J., Bogorad, L.: Two-dimensional polyacrylamide gel electrophoresis: an improved method for ribosomal proteins. Analyt. Biochem. 57, 200–210 (1974)

    Google Scholar 

  • Möller, W., Groene, A., Terhorst, C., Amons, R.: 50S ribosomal proteins. Purification and partial characterisation of two acidic proteins, A1 and A2, isolated from 50-S ribosomes of Escherichia coli. Europ. J. Biochem. 25, 5–12 (1972)

    Google Scholar 

  • Mira, G., Donner, D., Thammana, P., Lutter, L., Kurland, C. G.: Purification and characterisationof 50S ribosomal proteins of Escherichia coli. Molec. Gen. Genet. 112, 229–242 (1971)

    Google Scholar 

  • Nair, K. G., Arnstein, H. R. V.: Further observations on the polynucleotide-induced stimulation of protein synthesis by cell-free preparations from rabbit reticulocytes. Biochem. J. 97, 595–606 (1965)

    Google Scholar 

  • Reisfeld, R. A., Lewis, U. J., Williams, D. E.: Disc electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature (Lond.) 195, 281–283 (1962)

    Google Scholar 

  • Schweet, R., Lamfrom, H., Alden, E.: The synthesis of hemoglobin in a cell-free system. Proc. nat. Acad. Sci. (Wash.) 44, 1029–1035 (1958)

    Google Scholar 

  • Shapiro, A. L., Vinuela, E., Maizel, J. V., Jr.: Molecular weight estimation of polypeptide chains by electrophoresis on sodium dodecyl sulfate-polyacrylamide gels. Biochem. biophys. Res. Commun. 28, 815–820 (1967)

    Google Scholar 

  • Sherton, C. C., Wool, I. G.: Determination of the number of proteins in liver ribosomes and ribosomal subunits by two-dimensional polyacrylamide gel electrophoresis. J. biol. Chem. 247, 4460–4467 (1972)

    Google Scholar 

  • Sherton, C. C., Wool, I. G.: Two-dimensional polyacrylamide gel electrophoresis of eukaryotic ribosomal proteins. Methods in Enzymol. 30, 506–526 (1974)

    Google Scholar 

  • Spitnik-Elson, P.: The preparation of ribosomal protein from Escherichia coli with lithium cloride and urea. Biochem. biophys. Res. Commun. 18, 557–562 (1965)

    Google Scholar 

  • Stahl, J., Welfle, H., Bielka, H.: Studies on proteins of animal ribosomes. XIV. Analysis of phosphorylated rat liver ribosomal proteins by two-dimensional polyacrylamide gel electrophoresis. FEMS Letters 26, 233–236 (1972)

    Google Scholar 

  • Stöffler, G., Wool, I. G., Lin, A., Rak, K.-H.: The identification of the eukaryotic ribosomal proteins homologous with Escherichia coli proteins L7 and L12. Proc. nat. Acad. Sci. (Wash.) 71, 4723–4726 (1974)

    Google Scholar 

  • Subramanian, A. R.: Sensitive separations procedure for Escherichia coli ribosomal proteins and the resolution of high molecular weight components. Europ. J. Biochem. 45, 541–546 (1974)

    Google Scholar 

  • Terao, K., Ogata, K.: Characterisation of the proteins of the small subunits of rat liver ribosomes. Biochim. biophys. Acta (Amst.) 285, 473–482 (1972)

    Google Scholar 

  • Thammana, P., Kurland, C. G., Deusser, E., Weber, J., Maschler, R., Stöffler, G., Wittmann, H. G.: Structural and functional evidence for a repeated 50S subunit ribosomal protein. Nature (Lond.) New Biol. 242, 47–49 (1973)

    Google Scholar 

  • Welfle, H., Stahl, J., Bielka, H.: Studies on proteins of animal ribosomes. VIII. Two-dimensional polyacrylamide gel electrophoresis of ribosomal proteins of rat liver. Biochim. biophys. Acta (Amst.) 243, 416–419 (1971)

    Google Scholar 

  • Welfle, H., Stahl, J., Bielka, H.: Studies on proteins of animal ribosomes. XIII. Enumeration of ribosomal proteins of rat liver. FEBS Letters 26, 228–232 (1972)

    Google Scholar 

  • Westermann, P., Bielka, H.: Studies on proteins of animal ribosomes. XV. Proteins of the small subunit of rat liver ribosomes: isolation, anino acid composition, tryptic peptides and molecular weights. Molec. gen. Genet. 126, 349–356 (1973)

    Google Scholar 

  • Westermann, P., Bielka, H., Böttger, M.: Studies on proteins of animal ribosomes. VII. Isolation of proteins from rat liver ribosomes, their molecular weights, amino acid compositions and secondary structure. Molec. gen. Genet. 111, 224–234 (1971)

    Google Scholar 

  • Wettstein, F. O., Staehelin, T., Noll, H.: Ribosomal aggregate engaged in protein synthesis: characterization of the ergosome. Nature (Lond.) 197, 430–435 (1963)

    Google Scholar 

  • Wittmann, H. G., Stöffler, G., Hindennach, I., Kurland, C. G., Randall-Hazelbauer, L., Birge, E. A., Nomura, M., Kaltschmidt, E., Mizushima, S., Traut, R. R., Bickle, T. A.: Correlation of 30S ribosomal proteins of Escherichia coli isolated in different laboratories. Molec. gen. Genet. 111, 327–333 (1971)

    Google Scholar 

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Authors and Affiliations

  1. Department of Biophysics, King's College, London

    O. H. W. Martini & H. J. Gould

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  1. O. H. W. Martini
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  2. H. J. Gould
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Communicated by H. G. Wittmann

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Martini, O.H.W., Gould, H.J. Characterisation of eukaryotic ribosomal proteins. Molec. Gen. Genet. 142, 299–316 (1975). https://doi.org/10.1007/BF00271254

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  • DOI: https://doi.org/10.1007/BF00271254

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