Template-dependent polypeptide synthesis in a factor- and energy-free translation system promoted by pyridine

Itaru Nitta, Takuya Ueda, Takahiko Nojima, Kimitsuna Watanabe

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We demonstrate here that a high concentration (40-70%) of pyridine, an aromatic tertiary amine catalyst, is able to promote translation on ribosomes without the presence of soluble protein factors or chemical energy sources. Compared with Monro's fragment reaction [Methods Enzymol. 20, 472-481 (1971)], which reflects only the peptidyltransferase step, this novel translation system can produce polypeptides with chain lengths of at least several tens of residues depending on the template RNA. In the presence of 60% pyridine, poly(U) and poly(UC) promoted incorporation of the respective amino acids, phenylalanine and serine-leucine, twofold, whereas poly(A) promoted the incorporation of lysine by only 25%. The degrees of polymerization of phenylalanine and lysine were up to the decamer and around 40mer, respectively. In poly(UC)-dependent oligo(serine-leucine) synthesis, oligo-peptides with a serine and leucine alternate sequence were the main products. This novel pyridine system evidently differs from the non-enzymatic translation system reported by Gavrilova and Spirin [FEBS Lett. 17, 324-326 (1971)]; the former system displays partial resistance toward deproteinization reagents such as SDS and proteinase K, whereas the latter system is completely sensitive.

Original languageEnglish
Pages (from-to)841-849
Number of pages9
JournalJournal of biochemistry
Volume118
Issue number4
DOIs
Publication statusPublished - 1995 Jan 1
Externally publishedYes

Fingerprint

Leucine
Serine
Free energy
Phenylalanine
Peptides
Lysine
Peptidyl Transferases
Poly U
Endopeptidase K
Poly A
Ribosomes
Chain length
Polymerization
Amines
RNA
Amino Acids
Catalysts
pyridine
Proteins

Keywords

  • Escherichia coli
  • Pyridine
  • Ribosomal RNA
  • Thermits thermophilus
  • Translation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Template-dependent polypeptide synthesis in a factor- and energy-free translation system promoted by pyridine. / Nitta, Itaru; Ueda, Takuya; Nojima, Takahiko; Watanabe, Kimitsuna.

In: Journal of biochemistry, Vol. 118, No. 4, 01.01.1995, p. 841-849.

Research output: Contribution to journalArticle

Nitta, Itaru ; Ueda, Takuya ; Nojima, Takahiko ; Watanabe, Kimitsuna. / Template-dependent polypeptide synthesis in a factor- and energy-free translation system promoted by pyridine. In: Journal of biochemistry. 1995 ; Vol. 118, No. 4. pp. 841-849.
@article{b70bef55f436457d8bbde7e658b06810,
title = "Template-dependent polypeptide synthesis in a factor- and energy-free translation system promoted by pyridine",
abstract = "We demonstrate here that a high concentration (40-70{\%}) of pyridine, an aromatic tertiary amine catalyst, is able to promote translation on ribosomes without the presence of soluble protein factors or chemical energy sources. Compared with Monro's fragment reaction [Methods Enzymol. 20, 472-481 (1971)], which reflects only the peptidyltransferase step, this novel translation system can produce polypeptides with chain lengths of at least several tens of residues depending on the template RNA. In the presence of 60{\%} pyridine, poly(U) and poly(UC) promoted incorporation of the respective amino acids, phenylalanine and serine-leucine, twofold, whereas poly(A) promoted the incorporation of lysine by only 25{\%}. The degrees of polymerization of phenylalanine and lysine were up to the decamer and around 40mer, respectively. In poly(UC)-dependent oligo(serine-leucine) synthesis, oligo-peptides with a serine and leucine alternate sequence were the main products. This novel pyridine system evidently differs from the non-enzymatic translation system reported by Gavrilova and Spirin [FEBS Lett. 17, 324-326 (1971)]; the former system displays partial resistance toward deproteinization reagents such as SDS and proteinase K, whereas the latter system is completely sensitive.",
keywords = "Escherichia coli, Pyridine, Ribosomal RNA, Thermits thermophilus, Translation",
author = "Itaru Nitta and Takuya Ueda and Takahiko Nojima and Kimitsuna Watanabe",
year = "1995",
month = "1",
day = "1",
doi = "10.1093/oxfordjournals.jbchem.a124989",
language = "English",
volume = "118",
pages = "841--849",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "4",

}

TY - JOUR

T1 - Template-dependent polypeptide synthesis in a factor- and energy-free translation system promoted by pyridine

AU - Nitta, Itaru

AU - Ueda, Takuya

AU - Nojima, Takahiko

AU - Watanabe, Kimitsuna

PY - 1995/1/1

Y1 - 1995/1/1

N2 - We demonstrate here that a high concentration (40-70%) of pyridine, an aromatic tertiary amine catalyst, is able to promote translation on ribosomes without the presence of soluble protein factors or chemical energy sources. Compared with Monro's fragment reaction [Methods Enzymol. 20, 472-481 (1971)], which reflects only the peptidyltransferase step, this novel translation system can produce polypeptides with chain lengths of at least several tens of residues depending on the template RNA. In the presence of 60% pyridine, poly(U) and poly(UC) promoted incorporation of the respective amino acids, phenylalanine and serine-leucine, twofold, whereas poly(A) promoted the incorporation of lysine by only 25%. The degrees of polymerization of phenylalanine and lysine were up to the decamer and around 40mer, respectively. In poly(UC)-dependent oligo(serine-leucine) synthesis, oligo-peptides with a serine and leucine alternate sequence were the main products. This novel pyridine system evidently differs from the non-enzymatic translation system reported by Gavrilova and Spirin [FEBS Lett. 17, 324-326 (1971)]; the former system displays partial resistance toward deproteinization reagents such as SDS and proteinase K, whereas the latter system is completely sensitive.

AB - We demonstrate here that a high concentration (40-70%) of pyridine, an aromatic tertiary amine catalyst, is able to promote translation on ribosomes without the presence of soluble protein factors or chemical energy sources. Compared with Monro's fragment reaction [Methods Enzymol. 20, 472-481 (1971)], which reflects only the peptidyltransferase step, this novel translation system can produce polypeptides with chain lengths of at least several tens of residues depending on the template RNA. In the presence of 60% pyridine, poly(U) and poly(UC) promoted incorporation of the respective amino acids, phenylalanine and serine-leucine, twofold, whereas poly(A) promoted the incorporation of lysine by only 25%. The degrees of polymerization of phenylalanine and lysine were up to the decamer and around 40mer, respectively. In poly(UC)-dependent oligo(serine-leucine) synthesis, oligo-peptides with a serine and leucine alternate sequence were the main products. This novel pyridine system evidently differs from the non-enzymatic translation system reported by Gavrilova and Spirin [FEBS Lett. 17, 324-326 (1971)]; the former system displays partial resistance toward deproteinization reagents such as SDS and proteinase K, whereas the latter system is completely sensitive.

KW - Escherichia coli

KW - Pyridine

KW - Ribosomal RNA

KW - Thermits thermophilus

KW - Translation

UR - http://www.scopus.com/inward/record.url?scp=0028825754&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028825754&partnerID=8YFLogxK

U2 - 10.1093/oxfordjournals.jbchem.a124989

DO - 10.1093/oxfordjournals.jbchem.a124989

M3 - Article

C2 - 8576102

AN - SCOPUS:0028825754

VL - 118

SP - 841

EP - 849

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 4

ER -