Skip to main content
SpringerLink
Log in

Localization of ribosomal protein L27 at the peptidyl transferase centre of the 50 S subunit, as determined by immuno-electron microscopy

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

Summary

Protein L27 has been localized on the ribosomal surface by immuno-electron microscopy by using antibodies specific for Escherichia coli L27, and by reconstituting 50 S subunits from an E. coli mutant, which lacks protein L27, with the homologous protein from Bacillus subtilis and using antibodies specific for the B. subtilis protein. With both approaches, protein L27 has been located at the base of the central protuberance at the interface side of the 50 S particle and thus in proximity to the peptidyl transferase centre. The immuno-electron microscopic data also suggest that the interface region of the 50 S particle is not as flat as most of the proposed three-dimensional models suggest, but instead there is a significant depression.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Breitenreuter G, Lotti M, Stöffler-Meilicke M, Stöffler G (1984) Comparative electron microscopic study on the location of ribosomal proteins S3 and S7 on the surface of the E. coli 30 S subunit using monoclonal and conventional antibody. Mol Gen Genet 197:189–195

    Google Scholar 

  • Czernilofsky AP, Collatz EE, Stöffler G, Kuechler E (1974) Proteins at the tRNA binding site of Escherichia coli ribosomes. Proc Natl Acad Sci USA 71:230–234

    Google Scholar 

  • Dabbs ER (1983) A pair of Bacillus subtilis ribosomal proteins mapping outside the principal ribosomal protein cluster. J Bacteriol 156:966–969

    Google Scholar 

  • Dabbs ER, Ehrlich R, Hasenbank R, Schroeter B-H, Stöffler-Meilicke M, Stöffler G (1981) Mutants of Escherichia coli lacking ribosomal protein L1. J Mol Biol 149:553–578

    Google Scholar 

  • Dabbs ER, Hasenbank R, Kastner B, Rak K-H, Wartusch B, Stöffler G (1983) Immunological studies of Escherichia coli mutants lacking one or two ribosomal proteins. Mol Gen Genet 192:301–308

    Google Scholar 

  • Geisser M, Tischendorf GW, Stöffler G (1973) Comparative immunological and electrophoretical studies on ribosomal proteins of Bacillaceae. Mol Gen Genet 127:129–145

    Google Scholar 

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

    Google Scholar 

  • Hauptmann R, Czernilofsky AP, Voorma HO, Stöffler G, Kuechler E (1974) Identification of a protein at the ribosomal donor-site by affinity labeling. Biochem Biophys Res Commun 56:331–337

    Google Scholar 

  • Herold M, Nierhaus KH (1987) Incorporation of six additional proteins to complete the assembly map of the 50 S subunit from Escherichia coli ribosomes. J Biol Chem 262:8826–8833

    Google Scholar 

  • Higo K, Otaka E, Osawa S (1982) Purification and characterization of 30 S ribosomal protein from Bacillus subtilis: Correlation to Escherichia coli 30 S proteins. Mol Gen Genet 185:239–244

    Google Scholar 

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

    Google Scholar 

  • Hjelm H, Hjelm K, Sjöquist J (1972) Protein A from staphylococcus aureus. Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglobins. FEBS Lett. 28:73–76

    Google Scholar 

  • Hofbauer C, Stöffler G, Otaka E, Osawa S (1984) Homology between the ribosomal proteins of Escherichia coli and Bacillus subtilis. Hoppe Seyler's Z Physiol Chem 365:922

    Google Scholar 

  • Lake JA, Strycharz W (1981) Ribosomal proteins L1, L17 and L27 from Escherichia coli localized at single sites on the large subunit by immune electron microscopy. J Mol Biol 153:979–992

    Google Scholar 

  • Lotti M, Dabbs ER, Hasenbank R, Stöffler-Meilicke M, Stöffler G (1983) Characterisation of a mutant from Escherichia coli lacking protein L15 and localisation of protein L15 by immunoelectron microscopy. Mol Gen Genet 192:295–300

    Google Scholar 

  • Lührmann R (1975) Identifizierung der Proteine S1, S13 und S18 als Komponenten der 30 S ribosomalen Codon-Bindestelle. Ph D Thesis, Universität Münster

  • Lührmann R, Bald R, Stöffler-Meilicke M, Stöffler G (1981) Localization of the puromycin binding site on the large ribosomal subunit of Escherichia coli by immunoelectron microscopy. Proc Natl Acad Sci USA 78:7276–7280

    Google Scholar 

  • Olson HM, Nicholson AW, Cooperman BS, Glitz DG (1985) Localization of sites of photoaffinity labeling of the large subunit of Escherichia coli ribosomes by arylazide derivative puromycin. J Biol Chem 260:10326–10331

    Google Scholar 

  • Pellegrini M, Oen H, Eilat D, Cantor CR (1974) The mechanism of covalent reaction of bromacetyl-phenylalanyl-transfer RNA binding site of the Escherichia coli ribosome. J Mol Biol 88:809–829

    Google Scholar 

  • Radermacher M, Wagenknecht T, Verschoor A, Frank J (1987) Three-dimensional structure of the large ribosomal subunit from Escherichia coli. EMBO J 6:1107–1114

    Google Scholar 

  • Schaber E, Kastner B, Stöffler-Meilicke M, Stöffler G (1984) Decoration of ribosomal subunits from Escherichia coli with monovalent Fab fragment specific for proteins L7/L12. In: Proceedings of the 8th european congress on electron microscopy, Budapest, vol 2, pp 1555–1556

  • Sonenberg N, Wilchek M, Zamir A (1973) Mapping of Escherichia coli ribosomal components involved in peptidyl transferase activity. Proc Natl Acad Sci USA 70:1423–1426

    Google Scholar 

  • Sopori M, Pellegrini M, Lengyel P, Cantor CR (1974) Affinity labeling of Escherichia coli ribosomal proteins with an analog of the natural initiator tRNA. Biochemistry 13:5432–5439

    Google Scholar 

  • Stöffler G, Wittmann HG (1971) Ribosomal proteins. XXV. Immunological studies on Escherichia coli ribosomal proteins. J Mol Biol 62:407–409

    Google Scholar 

  • Stöffler G, Stöffler-Meilicke M (1984) Immunoelectron microscopy of ribosomes. Annu Rev Biophys Bioeng 13:303–330

    Google Scholar 

  • Stöffler G, Hasenbank R, Lütgehaus M, Maschler R, Morrison CA, Zeichhardt H, Garrett RA (1973) The accessibility of proteins of the Escherichia coli ribosomal subunit to antibody binding. Mol Gen Genet 127:89–110

    Google Scholar 

  • Stöffler G, Bald R, Lührmann R, Tischendorf G, Stöffler-Meilicke M (1980) Localization of the binding sites of thiostrepton, chloramphenicol and puromycin on the 50 S ribosomal subunit by immuno electron microscopy. In: Brederow P, de Priester W (eds) Electron microscopy 1980 — biology. Seventh european congress on electron microscopy Foundation, Leiden, vol 2, pp 566–567

  • Stöffler G, Lotti M, Noah M, Stöffler-Meilicke M (1984a) The localization of proteins L5 and L10 on the surface of 50 S subunits of Escherichia coli. In: Proceedings of the 8th european congress on electron microscopy, Budapest, vol 2, pp 1551–1552

  • Stöffler G, Noah M, Stöffler-Meilicke M, Dabbs ER (1984b) The localization of protein L19 on the surface of 50 S subunits of Escherichia coli aided by the use of mutants lacking protein L19. J Biol Chem 259:4521–4526

    Google Scholar 

  • Stöffler-Meilicke M, Noah M, Stöffler G (1983a) Location of eight ribosomal proteins on the surface of the 50 S subunit from Escherichia coli. Proc Natl Acad Sci USA 80:6780–6784

    Google Scholar 

  • Stöffler-Meilicke M, Epe B, Steinhäuser KG, Woolley P, Stöffler G (1983b) Immunoelectron microscopy of ribosomes carrying a fluorescence label in a defined position. FEBS Lett. 163:94–98

    Google Scholar 

  • Stöffler-Meilicke M, Epe B, Woolley P, Lotti M, Littlechild J, Stöffler G (1984) Location of protein S4 on the small ribosomal subunit of E. coli and B. stearothermophilus with protein- and hapten-specific antibodies. Mol Gen Genet 197:8–18

    Google Scholar 

  • Stöffler-Meilicke M, Dabbs ER, Albrecht-Ehrlich R, Stöffler G (1985) A mutant from Escherichia coli which lacks ribosomal proteins S17 and L29 used to localize these two proteins on the ribosomal surface. Eur J Biochem 150:485–490

    Google Scholar 

  • Stöffler-Meilicke M, Stöffler G (1987) The localization of ribosomal proteins on the surface of the ribosomal subunits from Escherichia coli by immuno electron microscopy. Methods Enzymol (in press)

  • Tate WP, Dognin MJ, Noah M, Stöffler-Meilicke M, Stöffler G (1984) The NH2-terminal domain of Escherichia coli ribosomal protein L11. Its three-dimensional location and its role in the binding of release factors 1 and 2. J Biol Chem 259:7317–7324

    Google Scholar 

  • Tischendorf GW, Stöffler G (1975) Localization of Escherichia coli ribosomal protein S4 on the surface of the 30 S ribosomal subunit by immuno electron microscopy. I. Distribution of antibody-binding sites as obtained with immunoglobulins and monovalent antibody fragments from various S4-specific antisera. Mol Gen Genet 142:193–208

    Google Scholar 

  • Tischendorf GW, Zeichhardt H, Stöffler G (1974) Location of proteins S5, S13 and S14 on the surface of the 30 S ribosomal subunit from Escherichia coli as determined by immune electron microscopy. Mol Gen Genet 134:209–223

    Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

  • Yonath A, Leonard KR, Wittmann HG (1987) A tunnel in the large ribosomal subunit revealed by three-dimensional image reconstruction. Science 236:813–816

    Google Scholar 

  • Zubke W, Stadler H, Ehrlich R, Stöffler G, Wittmann HG (1977) Improved electrophoretic and immunochemical techniques for the identification and characterization of mutant proteins, applied to ribosomal protein S8 in Escherichia coli mutants. Mol Gen Genet 158:129–139

    Google Scholar 

Download references

Author information

Author notes

  1. Marina Stöffler-Meilicke

    Present address: Institut für Mikrobiologie der Medizinischen Fakultät, Universität Innsbruck, Fritz-Pregl-Strasse 3, A-6020, Innsbruck, Austria

  2. Marina Lotti

    Present address: Dipartimento di Fisiologia e Biochimica Generali, Universita Degli Studi di milano, Via Celoria 26, I-20133, Milano, Italy

Authors and Affiliations

  1. Max-Planck-Institut für Molekulare Genetik, Ihnestraße 73, D-1000, Berlin 33

    Marina Lotti & Marina Stöffler-Meilicke

  2. Institut für Mikrobiologie der Medizinischen Fakultät, Universität Innsbruck, Fritz-Pregl-Strasse 3, A-6020, Innsbruck, Austria

    Georg Stöffler

Authors
  1. Marina Lotti
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marina Stöffler-Meilicke
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Georg Stöffler
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by E. Bautz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lotti, M., Stöffler-Meilicke, M. & Stöffler, G. Localization of ribosomal protein L27 at the peptidyl transferase centre of the 50 S subunit, as determined by immuno-electron microscopy. Mol Gen Genet 210, 498–503 (1987). https://doi.org/10.1007/BF00327203

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00327203

Key words

Search

Navigation