The identification of ribosomal proteins that occur at, or near, the subunit interface of the 30S and 50S subunits in the E. coli 70S ribosome was attempted by studying the effect of antibodies on the Mg++ dependent dissociation-association equilibrium of 70S ribosomes. Dissociated ribosomes were mixed with monovalent fragments of IgG antibodies (Fab's) specific for each ribosomal protein and then reassociated into intact 70S particles. Various degrees of inhibition of this reassociation were observed for proteins S9, S11, S12, S14, S20, L1, L6, L14, L15, L19, L20, L23, L26 and L27. A small amount of aggregation of 50S subunits was caused by IgG's specific for the proteins S9, S11, S12, S14 and S20 and purified 50S subunits. It was inferred that the presence of small amounts of these proteins on 50S subunits was compatible with their presence at the subunit interface. Finally, the capacity of proteins S11 and S12 to bind to 23S RNA was demonstrated.
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Chang, F. N.: Conformational changes in ribosomal subunits following dissociation of the E. coli 70S ribosomes. J. molec. Biol. 78, 563–568 (1973)
Garrett, R. A., Rak, K. H., Daya, L., Stöffler, G.: Specific protein binding sites on 16S rRNA of E. coli. Molec. gen. Genet. 114, 112–124 (1971)
Gesteland, R. F.: Isolation and characterisation of ribonuclease I mutants of E. coli. J. molec. Biol. 16, 67–84 (1966)
Highland, J. H., Ochsner, E., 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.) in press (1974)
Hindennach, I., Kaltschmidt, E., Wittmann, H.-G.: Isolation of proteins from 50S ribosomal subunits of Escherichia coli. Europ. J. Biochem. 23, 12–16 (1971b)
Hindennach, I., Stöffler, G., Wittmann, H.-G.: Isolation of the proteins from 30S ribosomal subunits of E. coli. Europ. J. Biochem. 23, 7–11 (1971a)
Kaltschmidt, E., Wittmann, H.-G.: Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Analyt. Biochem. 36, 401–412 (1970)
Kikuchi, A., Monier, R.: Association of subunits in purified preparations of E. coli ribosomes. FEBS Letters 11, 157–159 (1970)
Kurland, C. G.: Molecular characterisation of RNA from E. coli ribosomes. 1. Isolation and molecular weights. J. molec. Biol. 2, 83–91 (1960)
Lowry, O. H., Rosebrough, A. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin-Phenol reagent. J. biol. Chem. 193, 265–275 (1951)
Marsh, R., Parmeggiani, A.: Requirement of 30S proteins S5 and S9 for the ribosome-dependent GTPase activity of elongation factor G. Proc. nat. Acad. Sci. (Wash.) 70, 151–155 (1973)
Moore, P. B.: Reaction of N-ethyl maleimide with the ribosome of E. coli. J. molec. Biol. 60, 169–184 (1971)
Putnam, F. W., Tan, H., Lyn, L. T., Easley, D. W., Shusuke, M.: The cleavage of rabbit gamma-globulin by papain. J. biol. Chem. 237, 717–726 (1962)
Robinson, H. K., Wade, H. G.: The preparation of ribosomal ribonucleic acid from whole bacteria. Biochem. J. 106, 897–903 (1968)
Sarma, V. R., Silverton, E. W., Davies, D. R., Terry, W. D.: The three-dimensional structure at 6 Å resolution of a human γG1 immunoglobulin molecule. J. biol. Chem. 246, 3753–3759 (1971)
Spirin, A. S.: On the equilibrium of the association-dissociation reaction of ribosomal subparticles and on the existence of the so-called ‘60S intermediate’ (“swollen 70S”) during centrifugation of the equilibrium mixture. FEBS Letters 14, 349–353 (1971)
Stöffler, G.: Immunochemical studies on the ribosome. Cold Spring Harbour Meeting. Eds. Tissières, Nomura and Lengyel. 1974. In press
Stöffler, G., Daya, L., Rak, K. H., Garrett, R. A.: Ribosomal proteins XXVI. The number of specific protein sites on 16S and 23S RNA of E. coli. J. molec. Biol. 62, 411–414 (1971)
Stöffler, G., Hasenbank, R., Lütgehaus, M., Maschler, R., Morrison, C., Zeichhardt, H., Garrett, R. A.: The accessibility of the proteins on the 30S ribosomal subunit of E. coli to antibody binding. Molec. gen. Genet. 127, 89–110 (1973)
Stöffler, G., Wittmann, H.-G.: Sequence differences of E. coli 30S ribosomal proteins as determined by immunological methods. Proc. nat. Acad. Sci. (Wash.) 68, 2283–2287 (1971a)
Stöffler, G., Wittmann, H.-G.: Immunological studies on E. coli ribosomal proteins. J. molec. Biol. 62, 407–409 (1971b)
Tissières, A., Watson, J. D., Schlessinger, D., Hollingworth, B. R.: Ribonucleoprotein particles from E. coli. J. molec. Biol. 1, 221–233 (1959)
Zamir, A., Miskin, R., Elson, D.: Inactivation and reactivation of ribosomal subunits. Aminoacyl transfer RNA binding activity of the 30S subunit of E. coli. J. molec. Biol. 60, 347–364 (1971)
Zitomer, R. S., Flaks, J. G.: Magnesium dependence and equilibrium of the Escherichia coli ribosomal subunit association. J. molec. Biol. 71, 263–279 (1972)
Paper No. 84 on “Ribosomal Proteins”. Preceding paper is by Rahmsdorf et al., Molec. gen. Genet. 127, 259–271 (1973).
Communicated by E. Bautz
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Morrison, C.A., Garrett, R.A., Zeichhardt, H. et al. Proteins occurring at, or near, the subunit interface of E. coli ribosomes. Molec. Gen. Genet. 127, 359–368 (1973). https://doi.org/10.1007/BF00267106
- Ribosomal Protein
- Subunit Interface