Peptide environment of the peptidyl transferase center from Escherichia coli 70 S ribosomes as determined by thermoaffinity labeling with dihydrospiramycin

Tools

Item Type:	Article
Title:	Peptide environment of the peptidyl transferase center from Escherichia coli 70 S ribosomes as determined by thermoaffinity labeling with dihydrospiramycin
Creators Name:	Bischof, O., Urlaub, H., Kruft, V. and Wittmann-Liebold, B.
Abstract:	In an attempt to gain information about the peptidyl transferase center at the peptide level we cross-linked the spiramycin derivative dihydrospiramycin to its functional binding site in the 70 S ribosome of Escherichia coli. In this manner ribosomal proteins S12, S14, L17, L18, L27 and L35 were found specifically affinity-labeled. Proteolytic fragmentation of these proteins, separation by C18 reversed-phase high performance liquid chromatography of the peptide mixtures, and subsequent sequence analysis of labeled peptides revealed peptide regions at positions Ala1-Lys9 and Tyr116-Lys119 of S12, Leu47-Asp53 of protein S14, Ser6-Lys35 of protein L17, Ala57-Lys63 of protein L18, Ala5-Lys18 and Val66-Lys71 of protein L27, and Thr5-Lys11 of protein L35. This approach is a valuable tool to characterize the binding site of spiramycin as well as the peptidyl transferase center at the molecular level.
Keywords:	Affinity Labels, Amino Acid Sequence, Anti-Bacterial Agents, Competitive Binding, Escherichia Coli, Kinetics, Molecular Sequence Data, Peptide Fragments, Peptidyl Transferases, Ribosomal Proteins, Ribosomes, Spiramycin
Source:	Journal of Biological Chemistry
ISSN:	0021-9258
Publisher:	American Society for Biochemistry and Molecular Biology
Volume:	270
Number:	39
Page Range:	23060-23064
Date:	29 September 1995
Official Publication:	https://doi.org/10.1074/jbc.270.39.23060
PubMed:	View item in PubMed

Repository Staff Only: item control page