Amino acid substitutions in protein biosynthesis. Poly(A)-directed polyphenylalanine synthesis.

J. M. Pezzuto, Sidney Hecht

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The fidelity of protein biosynthesis in vitro was studied quantitatively in a well defined system that employed poly(A) as a message for the elaboration of phenylalanine-containing polypeptides from Escherichia coli phenylalanyl-tRNALys. Admixture of phenylalanyl-tRNALys and lysyl-tRNALys in three different ratios resulted in the efficient formation of peptides that contained the amino acids in the same ratios in which they had been utilized in the individual incubation mixtures. The incorporation was also shown to be codon-specific in a quantitative sense; the poly(A)-directed incorporation of [14C]phenylalanine from phenylalanyl-tRNALys was unaffected by [3H]phenylalanyl tRNAPhe, by arginyl-tRNA (one species of which responds to the codon triplet AGA) or by unfractionated E. coli tRNA. These findings suggest that the transfer of amino acids from (misacylated) tRNAs into polypeptides in vitro is as predicted by the adapter hypothesis and that such systems can operate with sufficient fidelity to permit the preparation of proteins having defined amino acid substitutions.

Original languageEnglish (US)
Pages (from-to)865-869
Number of pages5
JournalJournal of Biological Chemistry
Volume255
Issue number3
StatePublished - Feb 10 1980
Externally publishedYes

Fingerprint

RNA, Transfer, Lys
Poly A
Biosynthesis
Protein Biosynthesis
Amino Acid Substitution
Substitution reactions
Amino Acids
Transfer RNA
Phenylalanine
Codon
Escherichia coli
Peptides
Proteins
Amino Acid-Specific Transfer RNA
RNA, Transfer, Phe
polyphenylalanine

ASJC Scopus subject areas

  • Biochemistry

Cite this

Amino acid substitutions in protein biosynthesis. Poly(A)-directed polyphenylalanine synthesis. / Pezzuto, J. M.; Hecht, Sidney.

In: Journal of Biological Chemistry, Vol. 255, No. 3, 10.02.1980, p. 865-869.

Research output: Contribution to journalArticle

@article{aea87335882b49b7a26ff92e79eebda0,
title = "Amino acid substitutions in protein biosynthesis. Poly(A)-directed polyphenylalanine synthesis.",
abstract = "The fidelity of protein biosynthesis in vitro was studied quantitatively in a well defined system that employed poly(A) as a message for the elaboration of phenylalanine-containing polypeptides from Escherichia coli phenylalanyl-tRNALys. Admixture of phenylalanyl-tRNALys and lysyl-tRNALys in three different ratios resulted in the efficient formation of peptides that contained the amino acids in the same ratios in which they had been utilized in the individual incubation mixtures. The incorporation was also shown to be codon-specific in a quantitative sense; the poly(A)-directed incorporation of [14C]phenylalanine from phenylalanyl-tRNALys was unaffected by [3H]phenylalanyl tRNAPhe, by arginyl-tRNA (one species of which responds to the codon triplet AGA) or by unfractionated E. coli tRNA. These findings suggest that the transfer of amino acids from (misacylated) tRNAs into polypeptides in vitro is as predicted by the adapter hypothesis and that such systems can operate with sufficient fidelity to permit the preparation of proteins having defined amino acid substitutions.",
author = "Pezzuto, {J. M.} and Sidney Hecht",
year = "1980",
month = "2",
day = "10",
language = "English (US)",
volume = "255",
pages = "865--869",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "3",

}

TY - JOUR

T1 - Amino acid substitutions in protein biosynthesis. Poly(A)-directed polyphenylalanine synthesis.

AU - Pezzuto, J. M.

AU - Hecht, Sidney

PY - 1980/2/10

Y1 - 1980/2/10

N2 - The fidelity of protein biosynthesis in vitro was studied quantitatively in a well defined system that employed poly(A) as a message for the elaboration of phenylalanine-containing polypeptides from Escherichia coli phenylalanyl-tRNALys. Admixture of phenylalanyl-tRNALys and lysyl-tRNALys in three different ratios resulted in the efficient formation of peptides that contained the amino acids in the same ratios in which they had been utilized in the individual incubation mixtures. The incorporation was also shown to be codon-specific in a quantitative sense; the poly(A)-directed incorporation of [14C]phenylalanine from phenylalanyl-tRNALys was unaffected by [3H]phenylalanyl tRNAPhe, by arginyl-tRNA (one species of which responds to the codon triplet AGA) or by unfractionated E. coli tRNA. These findings suggest that the transfer of amino acids from (misacylated) tRNAs into polypeptides in vitro is as predicted by the adapter hypothesis and that such systems can operate with sufficient fidelity to permit the preparation of proteins having defined amino acid substitutions.

AB - The fidelity of protein biosynthesis in vitro was studied quantitatively in a well defined system that employed poly(A) as a message for the elaboration of phenylalanine-containing polypeptides from Escherichia coli phenylalanyl-tRNALys. Admixture of phenylalanyl-tRNALys and lysyl-tRNALys in three different ratios resulted in the efficient formation of peptides that contained the amino acids in the same ratios in which they had been utilized in the individual incubation mixtures. The incorporation was also shown to be codon-specific in a quantitative sense; the poly(A)-directed incorporation of [14C]phenylalanine from phenylalanyl-tRNALys was unaffected by [3H]phenylalanyl tRNAPhe, by arginyl-tRNA (one species of which responds to the codon triplet AGA) or by unfractionated E. coli tRNA. These findings suggest that the transfer of amino acids from (misacylated) tRNAs into polypeptides in vitro is as predicted by the adapter hypothesis and that such systems can operate with sufficient fidelity to permit the preparation of proteins having defined amino acid substitutions.

UR - http://www.scopus.com/inward/record.url?scp=0019320635&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0019320635&partnerID=8YFLogxK

M3 - Article

C2 - 6985907

AN - SCOPUS:0019320635

VL - 255

SP - 865

EP - 869

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 3

ER -