Summary
The accessibility of each of the proteins on the 50S ribosomal subunit of Escherichia coli was investigated by establishing whether immunoglobulins (IgG), specific for each of the 34 proteins from the 50S subunit, were able to bind to the 50S subunit. The main criterion for accessibility was the formation of specific antibody-50S subunit complexes that could be detected by means of analytical ultracentrifugation.
The proteins fell into two main groups. Immunoglobulins against proteins L1, L2, L3, L4, L5, L6, L7/L12, L8, L9, L10, L11, L14, L15, L16, L17, L18, L19, L20, L21, L22, L23, L25, L26, L27 and L30 gave large amounts of complex (20–100%) and, therefore, these proteins were considered to be accessible sible on the surface of the 50S ribosomal subunit. The antibodies against the remaining proteins L13, L24, L28, L29 and L31 to L34 produced small amounts of complexes (10–20%). Since their effects were unequivocably stronger than those obtained with IgG's from sera of non-immunized animals, the results indicate that these proteins are probably also accessible. Nonetheless, from the ultracentrifugation studies alone definite conclusions about the exposure of the latter group of proteins could not be drawn.
Similar content being viewed by others
References
Chang, F.N., Flaks, J.G.: Topography of the Escherichia coli ribosome. II. Preliminary sequence of 50S subunit protein attack by trypsin and its correlation with functional activities. J. molec. Biol. 61, 387–400 (1971)
Cotter, R.I., Gratzer, W.B.: Accessibility of RNA and protein in the ribosome. Investigation by hydrogen exchange and solvent perturbation. Europ. J. Biochem. 23, 468–474 (1971)
Crichton, R.R., Wittmann, H.G.: Trypsin digestion as a possible probe of the conformation of Escherichia coli ribosomes. Molec. gen. Genet. 114, 95–105 (1971)
Crichton, R.R., Wittmann, H.G.: A native ribonucleoprotein complex from Escherichia coli ribosomes. Proc. nat. Acad. Sci. (Wash.) 70, 665–668 (1973)
Highland, J.H., Gordon, J., Bodley, J., Hasenbank, R., Stöffler, G.: Inhibition of elongation factor G by antibodies specific for several ribosomal proteins. Proc. nat. Acad. Sci. (Wash.) 70, 147–150 (1973)
Highland, J.H., Ochsner, E., Bodley, J., Hasenbank, R., Stöffler, G.: Coordinate inhibition of elongation factor G function and ribosomal subunit association by antibodies to several ribosomal proteins. Proc. nat. Acad. Sci. (Wash.) 71, 627–630 (1974)
Hindennach, I., Kaltschmidt, E., Wittmann, H.G.: Isolation of proteins from 50S ribosomal subunits of E. coli. Europ. J. Biochem. 23, 7–11 (1971)
Kahan, L., Kaltschmidt, E.: Glutaraldehyde reactivity of E. coli ribosomes. Biochemistry 11, 2691–2698 (1972)
Kaltschmidt, E., Witmann, H.G.: Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Anal. Biochem. 36, 401–412 (1970)
Lake, J.A., Kahan, L.: Ribosomal proteins S5, S11, S13 and S19 localised by electron microscopy of antibody labelled subunits. J. molec. Biol. 99, 631–644 (1975)
Lake, J.A., Pendergast, M., Kahan, L., Nomura, M.: Localisation of Escherichia coli ribosomal proteins S4 and S14 by electron microscopy of antibody labelled subunits. Proc. nat. Acad. Sci. (Wash.) 71, 4688–4692 (1974)
Lelong, J.C., Gros, D., Gros, F., Bollen, A., Maschler, R., Stöffler, G.: Function of individual 30S subunit proteins of E. coli. The effect of specific immunoglobulin fragments (Fab) on the activities of ribosomal decoding sites. Proc. nat. Acad. Sci. (Wash.) 71, 248–252 (1974)
Litman, D.J., Cantor, C.: Surface topography of the Escherichia coli ribosome. Enzymatic iodination of the 50S subunit. Biochemistry 13, 512–518 (1974)
Maschler, R.: Immunochemische Untersuchungen zur Funktion der Einzelproteine der 30S-Untereinheit von Escherichia coli Ribosomen. PhD Thesis, Freie Universität, Berlin, 1973
Michalski, C.J., Sells, B.H.: Molecular morphology of ribosomes. Structural considerations of Escherichia coli ribosomal subunits utilising lactoperoxidase-catalysed iodination of ribosomal proteins. Europ. J. Biochem. 49, 361–367 (1974)
Möller, W., Groene, A., Terhorst, C., Amons, B.: 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)
Moore, G., Crichton, R.R.: Reductive methylation: A method for preparing functionally active radioactive ribosomes. FEBS Letters 37, 74–78 (1973)
Morrison, C.A.: Eine Studie zur Topographie von Proteinen von Escherichia coli Ribosomen mit proteinspezifischen Antikörpern und analytische Ultrazentrifugation. Ph.D. Thesis, Freie Universität, Berlin, 1974
Morrison, C.A., Garrett, R.A., Zeichhardt, H., Stöffler, G.: Proteins occurring at, or near, the subunit interface of E. coli ribosomes. Molec. gen. Genet. 127, 359–368 (1973)
Noll, M., Noll, H.: Structural dynamics of bacterial ribosomes. V. Magnesium-dependent dissociation of tight-couples into subunits: Measurements of dissociation constants and exchange rates. J. molec. Biol. 105, 111–130 (1976)
Schulte, C., Schiltz, E., Garrett, R.: The characterisation of a fragment of ribosomal protein S4 that is protected against trypsin digestion by 16S ribosomal RNA of Escherichia coli. Nucleic Acid Res. 2, 931–941 (1975)
Sonenberg, N., Wilchek, M., Zamir, A.: Mapping of Escherichia coli ribosomal components involved in peptidyl transferase activity. Proc. nat. Acad. Sci. (Wash.) 70, 1423–1426 (1973)
Spitnik-Elson, P., Breiman, A.: The effect of trypsin on 30S and 50S ribosomal subunits of Escherichia coli. Biochim. biophys. Acta (Amst.) 254, 457–467 (1971)
Stöffler, G.: A rapid micro-method for electrophoretic separation of Escherichia coli ribosomal proteins on gelatinised cellulose acetate. Molec. gen. Genet. 100, 374–377 (1967)
Stöffler, G.: Structure and function of the Escherichia coli ribosome: immunochemical analysis. In: Ribosomes (P. Lengyel, M. Nomura and A. Tissières, eds.). Long Island, New York. Cold Spring Harbor Laboratory 1974
Stöffler, G., Hasenbank, R., Lütgehaus, M., Maschler, R., Morrison, C.A., Zeichhardt, H., Garrett, R.A.: The accessibility of proteins of the Escherichia coli 30S ribosomal subunit to antibody binding. Molec. gen. Genet. 127, 89–110 (1973)
Stöffler, G., Tischendorf, G.W.: Antibiotic receptor sites in Escherichia coli ribosomes: In drug receptor interactions in antimicrobiol chemotherapy, Vol. 1. Topics in infectious diseases (J. Drews, F.E. Hahn, eds.). Wien-New York: Springer 1975
Stöffler, G., Wittmann, H.G.: Immunological studies on E. coli ribosomal proteins. J. molec. Biol. 62, 407–409 (1971a)
Stöffler, G., Wittmann, H.G.: Sequence difference of E. coli 30S ribosomal proteins as determined by immunolgical methods. Proc. nat. Acad. Sci. (Wash.) 68, 2283–2287 (1971b)
Stöffler, G., Wittmann, H.G.: Primary structure and threedimensional arrangement of proteins within the Escherichia coli ribosome (ed. H. Weissbach). In press, 1977
Tate, W.P., Caskey, C.T., Stöffler, G.: Inhibition of peptide chain termination by antibodies specific for ribosomal proteins. J. molec. Biol. 93, 375–389 (1975)
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.) 242, 47–49 (1973)
Tischendorf, G.W., Zeichhardt, H., Stöffler, G.: Determination of the location of proteins L14, L17, L18, L19, L22 and L23 on the surface of the 50S ribosomal subunit of Escherichia coli by immune electron microscopy. Molec. gen. Genet. 134, 187–202 (1974)
Tischendorf, G.W., Zeichhardt, H., Stöffler, G.: Architecture of the Escherichia coli ribosome as determined by immune electron microscopy. Proc. nat. Acad. Sci. (Wash.) 72, 4820–4824 (1975)
Visentin, L.P., Yaguchi, M., Kaplan, H.: Structure of the ribosome: Exposure of ribosomal proteins determined by competitive labelling. Canad. J. Biochem. 51, 1487–1497 (1973)
Wabl, M.R.: Electron microscopic localisation of two proteins on the surface of the 50S ribosomal subunit of Escherichia coli using specific antibody markers. J. molec. Biol. 84, 241–247 (1974)
Weber, H.J.: Stoichiometric measurements of 30S and 50S ribosomal proteins from Escherichia coli. Molec. gen. Genet. 119, 233–248 (1972)
Wittmann-Liebold, B., Dzionara, M.: Comparison of amino acid sequences among ribosomal proteins of Escherichia coli. FEBS Letters 61, 14–19 (1976a)
Wittmann-Liebold, B., Dzionara, M.: Studies on the significance of sequence homologies among proteins from Escherichia coli ribosomes. FEBS Letters 65, 281–283 (1976b)
Zeichhardt, H.: Immunochemische Untersuchungen zur Topographie der Proteine im Ribosom von Escherichia coli. Ph.D. Thesis. Freie Universität, Berlin 1976
Author information
Authors and Affiliations
Additional information
Communicated by E. Bautz
Rights and permissions
About this article
Cite this article
Morrison, C.A., Tischendorf, G., Stöffler, G. et al. Accessibility of proteins in 50S ribosomal subunits of Escherichia coli to antibodies: An ultracentrifugation study. Molec. Gen. Genet. 151, 245–252 (1977). https://doi.org/10.1007/BF00268787
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00268787