Identification of 50S proteins at the peptidyl-tRNA binding site of Escherichia coli ribosomes.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 70 ·No. 10 ·1973-10-00 ·Pages 2799-803

Oen H, Pellegrini M, Eilat D, Cantor CR

Abstract

Bromoacetyl-phenylalanyl-tRNA(phe) bound to 70S E. coli ribosomes reacts covalently with proteins of the 50S subunit. The major reactions are with proteins L2 and L27. In the presence of poly(U), 70S-bound bromoacetyl-phenylalanyl-tRNA(phe) can participate in peptidebond formation with phenylalanyl-tRNA(phe) or puromycin. Most of the products of these reactions are also found covalently attached to L2 and L27. Chloramphenicol and sparsomycin markedly inhibit the peptide-bond formation. These results strongly suggest that bromoacetylphenylalanyl-tRNA(phe) can function as a normal peptidyl-tRNA and that the 50S proteins, L2 and L27, are located in the peptidyl-tRNA binding site. The side reactions of bromoacetyl-phenylalanyl-tRNA(phe) are with one or more 50S proteins from the set L14-17, L6 and/or L11, and L26. These occur to a much less extent than the reactions with L2 and L27. Any functional significance of the side reactions is unknown.

MeSH Terms

Acetates Anti-Bacterial Agents/pharmacology Bacterial Proteins/isolation & purification Binding Sites Bromates Chloramphenicol/pharmacology Electrophoresis, Polyacrylamide Gel Escherichia coli/analysis Phenylalanine Protein Binding/drug effects Puromycin/pharmacology RNA, Transfer/metabolism Ribonucleosides/pharmacology Ribosomes/analysis Tritium

Chemicals

Acetates Anti-Bacterial Agents Bacterial Proteins Bromates Ribonucleosides Tritium Phenylalanine Puromycin Chloramphenicol RNA, Transfer

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Oen H

Pellegrini M

Eilat D

Cantor C R

References (16)

16 references, click to expand

A procedure for rapid and sensitive staining of protein fractionated by polyacrylamide gel electrophoresis.
Anal Biochem. 1967 Jul;20(1):150-4 PMID: 4166536
Structure and function of E. coli ribosomes. V. Reconstitution of functionally active 30S ribosomal particles from RNA and proteins.
Proc Natl Acad Sci U S A. 1968 Mar;59(3):777-84 PMID: 4868216
Ribosomal proteins of E. Coli: stoichiometry and implications for ribosome structure.
Cold Spring Harb Symp Quant Biol. 1969;34:25-38 PMID: 4985888
Ribosomal proteins. VII. Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins.
Anal Biochem. 1970 Aug;36(2):401-12 PMID: 4916449
Ribosomal proteins. 8. Molecular weights of isolated ribosomal proteins of Escherichia coli.
Proc Natl Acad Sci U S A. 1970 Dec;67(4):1909-13 PMID: 4923120
Studies on rat liver phenylalanyl transfer ribonucleic acid synthetase. I. Purification, stabilization, and complex formation.
J Biol Chem. 1971 Jun 10;246(11):3494-9 PMID: 5578904
Reaction of N-ethyl maleimide with the ribosomes of Escherichia coli.
J Mol Biol. 1971 Aug 28;60(1):169-84 PMID: 4937190
Ribosomal proteins. Isolation of proteins from 50S ribosomal subunits of Escherichia coli.
Eur J Biochem. 1971 Nov 11;23(1):12-6 PMID: 4942547
Ribosomal proteins. XXX. Specific protein binding sites on 23S RNA of Escherichia coli.
Mol Gen Genet. 1972;114(2):125-33 PMID: 4553097
Ribosomal proteins. XXIV. Trypsin digestion as a possible probe of the conformation of Escherichia coli ribosomes.
Mol Gen Genet. 1972;114(2):95-105 PMID: 4553100
Covalent attachment of a peptidyl-transfer RNA analog to the 50S subunit of Escherichia coli ribosomes.
Proc Natl Acad Sci U S A. 1972 Apr;69(4):837-41 PMID: 4554533
Phosphorylation of ribosomal proteins of Escherichia coli by protein kinase from rabbit skeletal muscle.
Biochemistry. 1972 Jun 20;11(13):2503-9 PMID: 4339244
Reconstitution of 50S ribosomal subunits from protein-free ribonucleic acid.
Biochemistry. 1973 Jan 16;12(2):220-4 PMID: 4682994
Stoichiometric measurements of 30S and 50S ribosomal proteins from Escherichia coli.
Mol Gen Genet. 1972;119(3):233-48 PMID: 4567157
Mapping of Escherichia coli ribosomal components involved in peptidyl transferase activity.
Proc Natl Acad Sci U S A. 1973 May;70(5):1423-6 PMID: 4576018
Reaction of celite-bound fluorescein isothiocyanate with the 50S E. coli ribosomal subunit.
Arch Biochem Biophys. 1973 Jul;157(1):125-32 PMID: 4352054

Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.70.10.2799

Published

1973-10-00

Pages

2799-803

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC427112

Subset

External Links

PubMed source DOI: 10.1073/pnas.70.10.2799 PMC Full Text View journal details

Analysis Services

Identification of 50S proteins at the peptidyl-tRNA binding site of Escherichia coli ribosomes.

Contact

WeChat Official Account