Location and domain structure of Escherichia coli ribosomal protein L7/L12: site specific cysteine cross-linking and attachment of fluorescent probes
10.1139/o95-102
Crossref journal-article
Canadian Science Publishing
Biochemistry and Cell Biology (155)
Abstract

Five different variants of L7/L12 containing single cysteine substitutions, two in the N-terminal (NTD) and three in the C-terminal domain (CTD), were produced, modified with [125I]N-[4-(p-azidosalicylamido)butyl]-3-(2′-pyridyldithio) propionamide ([125I]APDP), a sulfhydryl-specific, heterobifunctional, cleavable photo-cross-linking reagent, and reconstituted into ribosomes. These were irradiated, the total proteins were extracted and reductively cleaved, and the cross-linked proteins were identified. The effect of zero-length disulfide cross-linking on binding and activity was also determined. The same sites in L7/L12 were used to attach a rhodamine dye. The formation of ground-state rhodamine dimers caused the appearance of a new absorption band at 518 nm that was used to estimate the extent of interaction of the probes in the free protein and in complexes with L10. The three sites in the CTD, but not the N-terminal sites, cross-linked to L2 and L5 and to 30S proteins S2, S3, S7, S14, and S18 in a manner influenced by elongation factors. Binding to the ribosome and, therefore, function were blocked by zero-length cross-linking within the NTD, but not the CTD. Binding also disrupted rhodamine dimers in the NTD. No rhodamine dimers formed in the CTD.Key words: ribosomes; L7/L12; cross-links; rhodamine dimers.

Bibliography

Traut, R. R., Dey, D., Bochkariov, D. E., Oleinikov, A. V., Jokhadze, G. G., Hamman, B., & Jameson, D. (1995). Location and domain structure of Escherichia coli ribosomal protein L7/L12: site specific cysteine cross-linking and attachment of fluorescent probes. Biochemistry and Cell Biology, 73(11–12), 949–958.

Authors 7
  1. Robert R. Traut (first)
  2. Debendranath Dey (additional)
  3. Dmitry E. Bochkariov (additional)
  4. Andrew V. Oleinikov (additional)
  5. George G. Jokhadze (additional)
  6. Brian Hamman (additional)
  7. David Jameson (additional)
References 42 Referenced 52
  1. 10.1016/0006-291X(75)90818-9 / Biochem. Biophys. Res. Commun. by Agthoven A (1975)
  2. 10.1016/S0021-9258(18)83771-1 / J. Biol. Chem. by Bushuev V.N. (1989)
  3. 10.1016/S0021-9258(17)30576-8 / J. Biol. Chem. by Casiano C. (1990)
  4. 10.1016/S0021-9258(17)42543-9 / J. Biol. Chem. by Cowgill C.A. (1984)
  5. Falke, J.J., and Koshland, D.E.J. 1987. Global flexibility in a sensory receptor: a site-directed cross-linking approach. Science. (Washington, D.C.), 237: 1596-1600. (10.1126/science.2820061)
  6. 10.1016/0014-5793(81)80664-3 / FEBS Lett. by Girshovich AS. (1981)
  7. 10.1016/0014-5793(86)80325-8 / FEBS Lett. by Girshovich A. (1986)
  8. 10.1111/j.1432-1033.1978.tb12605.x / Eur. J. Biochem. by Gudkov A.T. (1978)
  9. 10.1016/0014-5793(82)80448-1 / FEBS Lett. by Gudkov A.T. (1982)
  10. 10.1016/S0021-9258(19)45679-2 / J. Biol. Chem. by Hamel E. (1972)
  11. {'key': 'p_11/p_11_1', 'first-page': '534550', 'volume': '59', 'author': 'Kenny J.W.', 'year': '1979', 'journal-title': 'Methods Enzymol.'} / Methods Enzymol. by Kenny J.W. (1979)
  12. Kischa, K., Moller, W., and Stoffler, G. 1971. Reconstitution of a GTPase activity by a 50s a ribosomal protein from E. coli. Nature, New Biol. 233: 6 2 4 3 . (10.1038/newbio233062a0)
  13. 10.1111/j.1432-1033.1978.tb12607.x / Eur. J. Biochem. by Koteliansky V.E. (1978)
  14. 10.1016/0022-2836(76)90200-X / J. Mol. Biol. by Lake J.A. (1976)
  15. 10.1016/0022-2836(86)90339-6 / J. Mol. Biol. by Langer J.A. (1986)
  16. 10.1016/0022-2836(87)90183-5 / J. Mol. Biol. by Leijonmarck M. (1987)
  17. Leijonmarck,M., Pettersson, I., and Liljas, A. 1981. Structural studies on the protein L7L12 from E. coli ribosomes. In Structural aspects of recognition and assembly of biological macromolecules. Edited by M. Balaban, J.L. Sussman, W. Traub, and A. Yonath. Balaban ISS, Rehovot and Philadelphia.pp. 761-776.
  18. 10.1016/0079-6107(82)90013-X / Prog. Biophys. Mol. Biol. by Liljas A. (1982)
  19. 10.1016/0300-9084(87)90004-6 / Biochimie by Liljas A. (1987)
  20. 10.1016/0300-9084(93)90146-J / Biochimie by Makarov E.M. (1993)
  21. Moller, W. 1990. Hypothesis: ribosomal protein L12 drives rotational movement of tRNA. In The ribosome: structure, function, and evolution. Edited by W.E. Hill, A. Dahlberg, R.A. Garrett, P.B. Moore, D. Schlessinger, and J.R. Warner. Am. Soc. Microbiol., Washington, D.C.
  22. Moller, W., and Maassen, J.A. 1986. On the structure, function, and dynamics of L7/L12 from Escherichia coli ribosomes. In Structure, function, and genetics of ribosomes. Edited by B. Hardesty and G. Kramer. Springer-Verlag,New York. pp. 309-325. (10.1007/978-1-4612-4884-2_18)
  23. 10.1016/0022-2836(83)90112-2 / J. Mol. Biol. by Moller W. (1983)
  24. 10.1073/pnas.90.21.9828 / Proc. Natl. Acad. Sci. U.S.A. by Oleinikov A.V. (1993)
  25. 10.1016/S0021-9258(18)54021-7 / J. Biol. Chem. by Oleinikov A.V. (1993)
  26. 10.1016/S0021-9258(19)62705-5 / J. Biol. Chem. by Olson H.M. (1986)
  27. 10.1111/j.1432-1033.1989.tb14731.x / Eur. J. Biochem. by Redl B. (1989)
  28. 10.1016/0300-9084(92)90108-Q / Biochimie by Scheinman A. (1992)
  29. 10.1016/S0021-9258(18)88812-3 / J. Biol. Chem. by Sommer A. (1985)
  30. Stoffler-Meilicke, M., and Stoffler, G. 1990. Topography of the ribosomal proteins from E. coli within the intact subunits as determined by immunoelectron microscopy and protein-protein crosslinking. In The ribosome: structure, function, and evolution. Edited by W.E. Hill, A. Dahlberg, R.A. Garrett, P.B. Moore, D. Schlessinger, and J.R. Warner. Am. Sci. Microbiol., Washington, D.C. pp. 123-133.
  31. 10.1016/0022-2836(78)90355-8 / J. Mol. Biol. by Strycharz W.A. (1978)
  32. 10.1111/j.1432-1033.1973.tb02740.x / Eur. J. Biochem. by Terhorst C. (1973)
  33. 10.1016/S0021-9258(19)62704-3 / J. Biol. Chem. by Tewari D.S. (1986)
  34. 10.1016/0022-2836(81)90250-3 / J. Mol. Biol. by Tokimatsu H. (1981)
  35. 10.1016/S0021-9258(17)43904-4 / J. Biol. Chem. by Traut R.R. (1983)
  36. Traut, R.R., Tewari, D.S., Sommer, A., Gavino, G., Olson, H.M., and Glitz, D.G. 1986. Protein topography of ribosomal functional domains: effects of monoclonal antibodies to different epitopes in Escherichia coli protein L7/L12 on ribosome function and structure. In Structure, function, and genetics of ribosomes. Edited by B. Hardesty and G.Kramer. Springer-Verlag, New York. pp. 286-308. (10.1007/978-1-4612-4884-2_17)
  37. Traut, R.R., Oleinikov, A.V., Makarov, E., Jokhadze, G., Perroud, B., and Wang, B. 1993. Structure and function of Escherichia coli ribosomal protein L7L12: effect of cross-links and deletions. In The translational apparatus: structure, function, regulation, evolution. Edited by K.H. Nierhaus, F. Franceschi, A.R. Subramanian, V.A. Erdmann, and B. Wittmann-Liebold. Plenum Press, New York. pp. 521-532. (10.1007/978-1-4615-2407-6_49)
  38. 10.1016/0006-291X(91)91566-U / Biochem. Biophys. Res. Commun. by Tsurugi K. (1991)
  39. 10.1073/pnas.84.16.5580 / Proc. Natl. Acad. Sci. U.S.A. by Uchiumi T. (1987)
  40. {'key': 'p_40/p_40_1', 'first-page': '189', 'volume': '92', 'author': 'Verschoor A.', 'year': '1986', 'journal-title': 'J. Ultrastruc. Res.'} / J. Ultrastruc. Res. by Verschoor A. (1986)
  41. 10.1021/bi00155a003 / Biochemistry by Zecherle G.N. (1992)
  42. 10.1016/S0021-9258(18)42637-3 / J. Biol. Chem. by Zecherle G.N. (1992)
Dates
Type When
Created 15 years, 8 months ago (Dec. 23, 2009, 2:51 p.m.)
Deposited 1 month, 3 weeks ago (July 2, 2025, 2:31 p.m.)
Indexed 1 month, 3 weeks ago (July 2, 2025, 3:10 p.m.)
Issued 29 years, 8 months ago (Dec. 1, 1995)
Published 29 years, 8 months ago (Dec. 1, 1995)
Published Print 29 years, 8 months ago (Dec. 1, 1995)
Funders 0

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@article{Traut_1995, title={Location and domain structure of Escherichia coli ribosomal protein L7/L12: site specific cysteine cross-linking and attachment of fluorescent probes}, volume={73}, ISSN={1208-6002}, url={http://dx.doi.org/10.1139/o95-102}, DOI={10.1139/o95-102}, number={11–12}, journal={Biochemistry and Cell Biology}, publisher={Canadian Science Publishing}, author={Traut, Robert R. and Dey, Debendranath and Bochkariov, Dmitry E. and Oleinikov, Andrew V. and Jokhadze, George G. and Hamman, Brian and Jameson, David}, year={1995}, month=dec, pages={949–958} }