Summary
Ribosomal proteins S1, S2, S16 and S23 were localized on the surface of the small subunit (40S) of rat liver ribosomes by immune electron microscopy. Antibodies against the single proteins were raised in rabbits and chicken and purified by affinity chromatography. 40S-IgG-40S complexes were obtained by incubation of 40S subunits with non-crossreacting antibodies specific for each of the four proteins and subsequent sucrose density gradient centrifugation. The location of the proteins was determined by means of antibody binding sites visualized in negative contrast in the electron microscope. The four investigated proteins are mainly located in the head region of the small subunit. Exposed antigenic determinants of proteins S1 and S2 were found to be located at different sites of the small subunit whereas proteins S16 and S23 were mapped in a limited region only.
Similar content being viewed by others
References
Amelunxen, F., Spiess, E.: Untersuchungen zur Struktur der Ribosomen. Ein Beitrag zur Konformation der 80S-Ribosomen von Pisum sativum. Cytobiologie 4, 293–306 (1971)
Bielka, H., Stahl, J.: Structure and function of eukaryotic ribosomes. Int. Rev. Biochem. 18, 79–168 (1978)
Blobel, G., Sabatini, D.: Dissociation of mammalian polyribosomes into subunits by puromycin. Proc. Natl. Acad. Sci. U.S.A. 68, 390–394 (1971)
Boublik, M., Hellmann, W., Kleinschmidt, A.K.: Size and structure of E. coli ribosomes by electron microscopy. Cytobiologie 14, 293–300 (1977)
Boublik, M., Hellmann, W., Roth, H.E.: Localization of ribosomal proteins L7/L12 in the 50S subunit of Escherichia coli ribosomes by electron microscopy. J. Mol. Biol. 107, 479–490 (1976)
Kiselev, N.A., Stel'maschuk, V.Ya., Lerman, M.I., Abakumova, O.Yu.: On the structure of liver ribosomes. J. Mol. Biol. 86, 577–586 (1974)
Lake, J.A.: Ribosome structure determined by electron microscopy of Escherichia coli small subunits, large subunits and monomeric ribosomes. J. Mol. Biol. 105, 131–159 (1976)
Lake, J.A., Kahan, L.: Ribosomal proteins S5, S11, S13 and S19 localized by electron microscopy of antibody-labeled subunits. J. Mol. Biol. 99, 631–644 (1975)
Lake, J.A., Pendergast, M., Kahan, L., Nomura, M.: Localization of Escherichia coli ribosomal proteins S4 and S14 by electron microscopy of antibody-labeled subunits. Proc. Natl. Acad. Sci. U.S.A. 71, 4688–4692 (1974)
Lutsch, G., Bielka, H., Wahn, K., Stahl, J.: Studies on the structure of animal ribosomes. III. Electron microscopic investigations of isolated rat liver ribosomes and their subunits Acta Biol. Med. Ger. 29, 851–876 (1972)
Lutsch, G., Noll, F., Theise, H., Bielka, H.: Localization of ribosomal protein S2 in rat liver ribosomes by immune electron microscopy. Acta Biol. Med. Ger. 36, 287–290 (1977)
Lutsch, G., Pleissner, K.-P., Wangermann, G., Noll, F.: Studies on the structure of animal ribosomes. VIII. Application of a digital image processing method to the enhancement of electron micrographs of small ribosomal subunits. Acta Biol. Med. Ger. 36, K59-K62 (1977)
McConkey, E.H., Bielka, H., Gordon, J., Lastick, S.M., Lin, A., Ogata, K., Reboud, J.-P., Traugh, J.A., Traut, R.R., Warner, J.R., Welfle, H., Wool, I.G.: Proposed uniform nomenclature for mammalian ribosomal proteins. Mol. Gen. Genet. 169, 1–6 (1979)
Noll, F., Bommer, U.A., Lutsch, G., Theise, H., Bielka, H.: Localization of rat liver ribosomal proterins S2 and its involvement in initiation factor eIF-2 binding to the 40S ribosomal subunit. FEBS Lett. 87, 129–131 (1978)
Noll, F., Theise, H., Bielka, H.: Studies on proteins of animal ribosomes. XVIII. Reaction of ribosomes and ribosomal proteins with antibodies against ribosomal proteins. Acta Biol. Med. Ger. 33, 547–553 (1974)
Noll, F., Wahn, K., Bielka, H.: Studies on the structure of animal ribosomes. IV. Electron microscopic projection of ribosome — antibody complexes. Acta Biol. Med. Ger. 31, 807–812 (1973)
Nonomura, Y., Blobel, G., Sabatini, D.: Structure of liver ribosomes studied by negative staining. J. Mol. Biol. 60, 303–323 (1971)
Politz, S.M., Glitz, D.G.: Ribosome structure Localization of N6, N6-dimethyladenosine by electron microscopy of a ribosome-antibody complex. Proc. Natl. Acad. Sci. U.S.A. 74, 1468–1472 (1977)
Reichelt, R., König, T., Wangermann, G.: Preparation of microgrids as specimen supports for high resolution electron microscopy. Micron 8, 29–31 (1977)
Stöffler, G.: Structure and function of the Escherichia coli ribosome: Immunochemical analysis. In: Ribosomes (M. Nomura, A. Tissières, P. Lengyel, eds.), pp. 615–667. Cold Spring Harbor Lab. 1974
Stöffler, G., Wittmann, H.G.: Primary structure and threedimensional arrangement of proteins within the Escherichia coli ribosome. In: Molecular mechanisms of protein biosynthesis (H. Weissbach and S. Pestka, (eds.), pp. 177–202. New York: Acad. Press 1977
Theise, H., Noll, F., Bielka, H.: Studies on proteins of animal ribosomes. XXVIII. Preparation and antigenic properties of 40S subunit proteins of rat liver ribosomes. Acta. Biol. Med. Ger. 37, 1353–1362 (1978)
Tischendorf, G.W., Stöffler, G.: Localization of Escherichia coli ribosomal protein S4 on the surface of the 30S ribosomal subunit by immune electron microscopy. Mol. Gen. Genet. 142, 193–208 (1975)
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. Mol. Gen. Genet. 134, 187–208 (1974a)
Tischendorf, G.W., Zeichhardt, H., Stöffler G.: Location of proteins S5, S13 and S14 on the surface of the 30S ribosomal subunits from Escherichia coli as determined by immune electron microscopy. Mol. Gen. Genet. 134, 209–223 (1974b)
Tischendorf, G.W., Zeichhardt, H., Stöffler, G.: Alchitecture of the Escherichia coli ribosomes as determined by immune electron microscopy. Proc. Natl. Acad. Sci. U.S.A. 72, 4820–4824 (1975)
Valentine, R.C., Shapiro, B.M., Stadtman, E.R.: Regulation of glutamine synthetase. XII. Electron microscopy, of the enzyme from Escherichia coli. Biochemistry 7, 2143–2152 (1968)
Vasiliev, V.D.: Morphology of the ribosomal 30S subparticle according to electron microscopic data. Acta Biol. Med. Ger. 33, 779–793 (1974)
Wabl, M.R.: Electron microscopic localization of two proteins on the surface of the 50S ribosomal subunit of Escherichia coli using specific antibody markers. J. Mol. Biol. 84, 241–247 (1974)
Welfle, H., Stahl, J., Bielka, H.: Studies on proteins of animal ribosomes. XII. Enumeration of ribosomal proteins of rat liver. FEBS Lett. 26, 228–230 (1972)
Westermann P., Heumann, W., Bielka, H.: On the stoichiometry of proteins in the small ribosomal subunit of hepatoma ascites cells. FEBS Lett. 62, 132–135 (1976)
Westermann, P., Heumann, W., Bommer, U.A., Bielka, H., Nygard, O., Hultin, T.: Cross-linking of initiation factor eIF-2 to proteins of the small subunit of rat liver ribosomes. FEBS Lett. 97, 101–104 (1979)
Wittmann, H.G.: Structure, function and evolution of ribosomes. Eur. J. Biochem. 61, 1–13 (1976)
Author information
Authors and Affiliations
Additional information
Communicated by H.G. Wittmann
S2,S3,S17,S21 according to the new nomenclature (McConkey et al., 1979)
Rights and permissions
About this article
Cite this article
Lutsch, G., Noll, F., Theise, H. et al. Localization of proteins S1, S2, S16 and S23 on the surface of small subunits of rat liver ribosomes by immune electron microscopy. Molec. Gen. Genet. 176, 281–291 (1979). https://doi.org/10.1007/BF00273223
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00273223