Preparation of misacylated aminoacyl-tRNA(Phe)'s useful as probes of the ribosomal acceptor site
| Autor: | Christopher K. Surratt, Cheng Xu, James R. Roesser, Robert C. Payne, Sidney M. Hecht |
|---|---|
| Rok vydání: | 1989 |
| Předmět: |
chemistry.chemical_classification
Peptide Biosynthesis Binding Sites Acylation Hydrolysis Pyroglutamyl-Peptidase I RNA Ligase (ATP) RNA Probes Ribosomal RNA Biology RNA Transfer Amino Acyl Biochemistry Ribosome Aminopeptidase Amino acid Pyroglutamate aminopeptidase chemistry Transfer RNA Escherichia coli Nucleotide T-Phages Ribosomes RNA ligase |
| Zdroj: | Biochemistry. 28(12) |
| ISSN: | 0006-2960 |
| Popis: | Several pyroglutamylaminoacyl-tRNA's were prepared by T4 RNA ligase mediated condensation of synthetic pyroglutamylaminoacyl-pCpA's with tRNA's from which the last two nucleotides at the 3'-end had been removed. The derived pyroglutamylaminoacyl-tRNA's were incubated in the presence of calf liver pyroglutamate aminopeptidase, which effected their conversion to free aminoacyl-tRNA's. The lack of contaminating esterase activities in the pyroglutamate aminopeptidase was verified by direct assay for the presence of the aminoacyl moieties in the formed aminoacyl-tRNA's and by the use of the deblocked aminoacyl-tRNA's as acceptors in the peptidyltransferase reaction using an Escherichia coli ribosomal system. These findings provide the wherewithal for a detailed investigation of the substrate specificity of the peptidyltransferase center and for the elaboration of polypeptides containing modified amino acids at predetermined sites. |
| Databáze: | OpenAIRE |
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