Preparation of misacylated aminoacyl-tRNAPhe's useful as probes of the ribosomal acceptor site

James R. Roesser, Cheng Xu, Robert C. Payne, Christopher K. Surratt, Sidney Hecht

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)5185-5195
Number of pages11
JournalBiochemistry
Volume28
Issue number12
StatePublished - 1989
Externally publishedYes

Fingerprint

RNA, Transfer, Phe
Transfer RNA
Pyroglutamyl-Peptidase I
Peptidyl Transferases
Esterases
Ligases
Substrate Specificity
Liver
Escherichia coli
Condensation
Assays
Nucleotides
RNA
Amino Acids
Peptides
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Roesser, J. R., Xu, C., Payne, R. C., Surratt, C. K., & Hecht, S. (1989). Preparation of misacylated aminoacyl-tRNAPhe's useful as probes of the ribosomal acceptor site. Biochemistry, 28(12), 5185-5195.

Preparation of misacylated aminoacyl-tRNAPhe's useful as probes of the ribosomal acceptor site. / Roesser, James R.; Xu, Cheng; Payne, Robert C.; Surratt, Christopher K.; Hecht, Sidney.

In: Biochemistry, Vol. 28, No. 12, 1989, p. 5185-5195.

Research output: Contribution to journalArticle

Roesser, JR, Xu, C, Payne, RC, Surratt, CK & Hecht, S 1989, 'Preparation of misacylated aminoacyl-tRNAPhe's useful as probes of the ribosomal acceptor site', Biochemistry, vol. 28, no. 12, pp. 5185-5195.
Roesser, James R. ; Xu, Cheng ; Payne, Robert C. ; Surratt, Christopher K. ; Hecht, Sidney. / Preparation of misacylated aminoacyl-tRNAPhe's useful as probes of the ribosomal acceptor site. In: Biochemistry. 1989 ; Vol. 28, No. 12. pp. 5185-5195.
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