Skip to main content
Log in

Ribosomal proteins

XXIV. Trypsin digestion as a possible probe of the conformation of Escherichia coli ribosomes

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

The susceptibility of 30S and 50S ribosomal subunits and of intact 70S ribosomes from E. coli to digestion with trypsin has been analysed by 2-dimensional electrophoresis. A classification of the ribosomal proteins on the basis of their rate of digestion by trypsin is proposed, and it is shown that it is possible to remove proteins by this method in a stepwise manner. The suitability of this method as a probe of the ribosomal conformation is discussed and compared with results from other investigations.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Chang, F. N., Flaks, J. G.: Topography of the Escherichia coli 30S ribosomal subunit and streptomycin binding. Proc. nat. Acad. Sci. (Wash.) 67, 1321–1328 (1970).

    Google Scholar 

  • Craven, G. R., Gupta, V.: Three-dimensional organisation of the 30S ribosomal proteins from Escherichia coli. I. Preliminary classification of the proteins. Proc. nat. Acad. Sci. (Wash.) 67, 1329–1336 (1970).

    Google Scholar 

  • —, Voynow, P., Hardy, S. J. S., Kurland, C. G.: Ribosomal proteins of E. coli. II. Chemical and physical characterisation of 30S proteins. Biochemistry (Wash.) 8, 2906–2915 (1969).

    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 

  • Fogel, S., Sypherd, P. S.: Chemical basis for heterogeneity of ribosomal proteins. Proc. nat. Acad. Sci. (Wash.) 59, 1329–1336 (1968).

    Google Scholar 

  • Garrett, R. A., Rak, K. H., Daya, L., Stöffler, G.: Ribosomal proteins, XXIX. Specific protein binding sites on 16S RNA of Escherichia coli. Molec. gen. Genet., submitted (1971).

  • Gesteland, R. F.: Isolation and characterisation of ribonuclease I mutants of Escherichia coli. J. molec. Biol. 16, 67–84 (1966).

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • —, Stöffler, G., Wittmann, H. G.: Ribosomal proteins. IX. Isolation of proteins from 30S ribosomal subunits of E. coli. Europ. J. Biochem. 23, 7–11 (1971a).

    Google Scholar 

  • Homann, H. E., Nierhaus, K. H.: Ribosomal proteins. Protein compositions of biosynthesic precursors and artifical subparticles from ribosomal subunits in Escherichia coli K12. Europ. J. Biochem. 20, 249–257 (1971).

    Google Scholar 

  • Kaltschmidt, E.: Ribosomal proteins. XIV. Isoelectric points of ribosomal proteins of E. coli as determined by two-dimensional polyacrylamide gel electrophoresis. Analyt. Biochem. 43, 25–31 (1971).

    Google Scholar 

  • Kaltschmidt, E., Dzionara, M., Donner, D., Wittmann, H. G.: Ribosomal proteins. I. Isolation, amino acid composition, molecular weights and peptide mapping of proteins from E. coli ribosomes. Molec. gen. Genet. 100, 364–373 (1967).

    Google Scholar 

  • —, Dzionara, M., Wittmann, H. G.: Ribosomal Proteins XV. Amino acid compositions of isolated ribosomal proteins from 30S and 50S Subunits of Escherichia coli. Molec. Gen. Genetics, 109, 292–297 (1970).

    Google Scholar 

  • —, Wittmann, H. G.: Ribosomal proteins. VII. Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Analyt. Biochem. 36, 401–412 (1970a).

    Google Scholar 

  • —: 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 

  • Mizushima, S., Nomura, M.: Assembly mapping of 30S ribosomal proteins from E. coli. Nature (Lond.) 226, 1214–1218 (1970).

    Google Scholar 

  • Moore, P. B., Traut, R. R., Noller, M., Pearson, P., Delius, H.: Ribosomal proteins of Escherrichia coli. II. Proteins from the 30S subunit. J. molec. Biol. 31, 441–461 (1968).

    Google Scholar 

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

    Google Scholar 

  • Nomura, M., Ozaki, M., Traub, P., Lowry, C. V.: Structure and function of ribosomes and their molecular components. Cold Spri. Harb. Symp. quant. Biol. 34, 49–61 (1969).

    Google Scholar 

  • Östner, U., Hultin, T.: The use of proteolytic enzymes in the study of ribosomal structure. Biochim. biophys. Acta (Amst.) 154, 376–387 (1968).

    Google Scholar 

  • Salas, M., Hille, M. B., Last, J. A., Wahba, A. J., Ochoa, S.: Translation of the genetic message. II. Effect of initiation factors on the binding of formyl-methionyl tRNA to ribosomes. Proc. nat. Acad. Sci. (Wash.) 57, 387–394 (1967).

    Google Scholar 

  • Schaup, H. W., Green, M., Kurland, C. G.: Molecular interactions of ribosomal components. I. Identification of RNA binding sites for individual 30S ribosomal proteins. Molec. gen. Genet. 109, 193–205 (1970).

    Google Scholar 

  • —: Molecular interactions of ribosomal components. II. Site-specific complex formation between 30S proteins and ribosomal RNA. Molec. gen. Genet. 112, 1–8 (1971).

    Google Scholar 

  • Stöffler, G., Daya, L., Rak, K. H., Garrett, R. A.: The number of specific protein binding sites on 16S and 23S RNA of Escherichia coli. J. molec. Biol., in press (1971a).

  • Stöffler, G., Daya, L., Rak, K. H., Garrett, R. A.: Specific protein binding sites on 23S RNA of Escherichia coli. Molec. gen. Genet., submitted (1971b).

  • —, Wittmann, H. G.: Sequence differences of Escherichia coli 30S ribosomal proteins as determined by immunochemical methods. Proc. nat. Acad. Sci. (Wash.) 68, 2283–2287 (1971).

    Google Scholar 

  • Traut, R. R., Delius, H., Ahmad-Zadek, C., Bickle, T. A., Pearson, P., Tissieres, A.: Ribosomal proteins of E. coli. Stoichiometry and implications for ribosome structure. Cold Spr. Harb. Symp. quant. Biol. 34, 25–38 (1969).

    Google Scholar 

  • —, Moore P. B., Delius, H., Noller, H., Tissieres, A.: Ribosomal proteins of E. coli. I. Demonstration of different primary structures. Proc. nat. Acad. Sci. (Wash.) 57, 1294–1301 (1967).

    Google Scholar 

  • Voynow, P., Kurland, C. G.: Stoichiometry of the 30S proteins of Escherichia coli. Biochemistry 10, 517–524 (1971).

    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 isolated in different laboratories. Molec. gen. Genet. 111, 327–333 (1971).

    Google Scholar 

  • Traut, R. R., Wittmann, H. G., Kaltschmidt, E., Rudloff, V., Janda, H. G., Dzionara, M., Donner, D., Nierhaus, K., Cech, M., Hindennach, I., Wittmann, B.: Protein-chemical and serological studies on ribosomes of bacteria, yeast and plants. FEBS Symposium, Madrid 1969, vol. 21, p. 33–46.

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by P. Starlinger

Rights and permissions

Reprints and permissions

About this article

Cite this article

Crichton, R.R., Wittmann, H.G. Ribosomal proteins. Molec. Gen. Genetics 114, 95–105 (1972). https://doi.org/10.1007/BF00332780

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00332780

Keywords

Navigation