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. 1973 Jul;70(7):1931–1935. doi: 10.1073/pnas.70.7.1931

A Protein Involved in the Peptidyltransferase Activity of Escherichia coli Ribosomes*

Knud H Nierhaus †,‡, Vicente Montejo §
PMCID: PMC433635  PMID: 4579005

Abstract

Cores were prepared from 50S ribosomal subunits by incubation with 0.4 M LiCl/Mg++ (0.4c cores); 0.8c cores and corresponding SP0.4-0.8 split proteins were obtained from 0.4c cores. In the fragment reaction 0.4c cores were active, but 0.8c cores were not. Activity of the 0.8c cores could be restored by reconstitution with the SP0.4-0.8 fraction. The split proteins were separated by DEAE-cellulose chromatography and Sephadex gel filtration. The peptidyltransferase activity is correlated with the amount of protein L11 added to the 0.8c core under reconstitution conditions. Whether protein L11 displays the enzymatic activity itself or is part of the enzymatic center is discussed.

Keywords: LiCl 50S core, ribosomal split proteins, reconstitution, peptide-bond formation

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Celma M. L., Monro R. E., Vazquez D. Substrate and antibiotic binding sites at the peptidyl transferase centre of E. coli ribosomes. FEBS Lett. 1970 Feb 16;6(3):273–277. doi: 10.1016/0014-5793(70)80076-x. [DOI] [PubMed] [Google Scholar]
  2. Celma M. L., Monro R. E., Vazquez D. Substrate and antibiotic binding sites at the peptidyl transferase centre of E. coli ribosomes: Binding of UACCA-Leu to 50 S subunits. FEBS Lett. 1971 Mar 16;13(4):247–251. doi: 10.1016/0014-5793(71)80546-x. [DOI] [PubMed] [Google Scholar]
  3. Fernandez-Munoz R., Monro R. E., Torres-Pinedo R., Vazquez D. Substrate- and antibiotic-binding sites at the peptidyl-transferase centre of Escherichia coli ribosomes. Studies on the chloramphenicol. lincomycin and erythromycin sites. Eur J Biochem. 1971 Nov 11;23(1):185–193. doi: 10.1111/j.1432-1033.1971.tb01607.x. [DOI] [PubMed] [Google Scholar]
  4. Gesteland R. F. Isolation and characterization of ribonuclease I mutants of Escherichia coli. J Mol Biol. 1966 Mar;16(1):67–84. doi: 10.1016/s0022-2836(66)80263-2. [DOI] [PubMed] [Google Scholar]
  5. Ghosh H. P., Söll D., Khorana H. G. Studies on polynucleotides. LXVII. Initiation of protein synthesis in vitro as studied by using ribopolynucleotides with repeating nucleotide sequences as messengers. J Mol Biol. 1967 Apr 28;25(2):275–298. doi: 10.1016/0022-2836(67)90142-8. [DOI] [PubMed] [Google Scholar]
  6. Haenni A. L., Chapeville F. The behaviour of acetylphenylalanyl soluble ribonucleic acid in polyphenylalanine synthesis. Biochim Biophys Acta. 1966 Jan 18;114(1):135–148. doi: 10.1016/0005-2787(66)90261-9. [DOI] [PubMed] [Google Scholar]
  7. Hindennach I., Kaltschmidt E., Wittmann H. G. Ribosomal proteins. Isolation of proteins from 50S ribosomal subunits of Escherichia coli. Eur J Biochem. 1971 Nov 11;23(1):12–16. doi: 10.1111/j.1432-1033.1971.tb01585.x. [DOI] [PubMed] [Google Scholar]
  8. Homann H. E., Nierhaus K. H. Ribosomal proteins. Protein compositions of biosynthetic precursors and artifical subparticles from ribosomal subunits in Escherichia coli K 12. Eur J Biochem. 1971 May 28;20(2):249–257. doi: 10.1111/j.1432-1033.1971.tb01388.x. [DOI] [PubMed] [Google Scholar]
  9. Kaltschmidt E. Ribosomal proteins. XIV. Isoelectric points of ribosomal proteins of E. coli as determined by two-dimensional polyacrylamide gel electrophoresis. Anal Biochem. 1971 Sep;43(1):25–31. doi: 10.1016/0003-2697(71)90103-5. [DOI] [PubMed] [Google Scholar]
  10. Kaltschmidt E., Wittmann H. G. Ribosomal proteins. VII. Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Anal Biochem. 1970 Aug;36(2):401–412. doi: 10.1016/0003-2697(70)90376-3. [DOI] [PubMed] [Google Scholar]
  11. Maden B. E., Traut R. R., Monro R. E. Ribosome-catalysed peptidyl transfer: the polyphenylalanine system. J Mol Biol. 1968 Jul 28;35(2):333–345. doi: 10.1016/s0022-2836(68)80028-2. [DOI] [PubMed] [Google Scholar]
  12. Monro R. E. Catalysis of peptide bond formation by 50 S ribosomal subunits from Escherichia coli. J Mol Biol. 1967 May 28;26(1):147–151. doi: 10.1016/0022-2836(67)90271-9. [DOI] [PubMed] [Google Scholar]
  13. Monro R. E., Cerná J., Marcker K. A. Ribosome-catalyzed peptidyl transfer: substrate specificity at the P-site. Proc Natl Acad Sci U S A. 1968 Nov;61(3):1042–1049. doi: 10.1073/pnas.61.3.1042. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Monro R. E., Staehelin T., Celma M. L., Vazquez D. The peptidyl transferase activity of ribosomes. Cold Spring Harb Symp Quant Biol. 1969;34:357–368. doi: 10.1101/sqb.1969.034.01.042. [DOI] [PubMed] [Google Scholar]
  15. Nierhaus K. H., Bordasch K., Homann H. E. Ribosomal proteins. 43. In vivo assembly of Escherichia coli ribosomal proteins. J Mol Biol. 1973 Mar 15;74(4):587–597. doi: 10.1016/0022-2836(73)90049-1. [DOI] [PubMed] [Google Scholar]
  16. Pestka S. Studies on the formation of transfer ribonucleic acid-ribosome complexes. X. Phenylalanyl-oligonucleotide binding to ribosomes and the mechanism of chloramphenicol action. Biochem Biophys Res Commun. 1969 Aug 15;36(4):589–595. doi: 10.1016/0006-291x(69)90345-3. [DOI] [PubMed] [Google Scholar]
  17. Siddiqui M. A., Hosokawa K. Role of 5S ribosomal RNA in polypeptide synthesis. II. Dissociation of 5S ribosomal RNA from 50S ribosomes in Escherichia coli. Biochem Biophys Res Commun. 1968 Jul 11;32(1):1–8. doi: 10.1016/0006-291x(68)90417-8. [DOI] [PubMed] [Google Scholar]
  18. Staehelin T., Maglott D. M., Monro R. E. On the catalytic center of peptidyl transfer: a part of the 50 S ribosome structure. Cold Spring Harb Symp Quant Biol. 1969;34:39–48. doi: 10.1101/sqb.1969.034.01.008. [DOI] [PubMed] [Google Scholar]
  19. Weber H. J. Stoichiometric measurements of 30S and 50S ribosomal proteins from Escherichia coli. Mol Gen Genet. 1972;119(3):233–248. doi: 10.1007/BF00333861. [DOI] [PubMed] [Google Scholar]

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