A novel cell-free system for peptide synthesis driven by pyridine

Itaru Nitta, Hirohide Nambu, Takaaki Okado, Shigeo Yoshinari, Takuya Ueda, Yaeta Endo, Knud H. Nierhaus, Kimitsuna Watanabe*


研究成果: Article査読

1 被引用数 (Scopus)


Previously we demonstrated that ribosomes can synthesize polypeptides in the presence of high concentrations (40-60%) of pyridine without any protein factors. Here we analyze additional ribosomal parameters in 60% pyridine using Escherichia coli ribosomes. Ribosomal subunits once exposed to pyridine failed to re-associate to 70S ribosomes in aqueous buffer systems even in the presence of 20 mM Mg2+, whereas they formed 70S complexes in the presence of 60% pyridine. Two-dimensional gel electrophoresis of ribosomal proteins revealed that some proteins located at the protuberances of the large subunit, e.g. L7/L12 and L11 forming the elongation factor-binding domain, were released in the pyridine system. The aminoglycoside neomycin, a strong inhibitor of the ribosomal (factor-independent) translocation reaction, completely blocked poly(Phe) synthesis and translocation activities in the pyridine system, whereas these activities were not affected at all by gypsophilin, a ribotoxin that inhibits factor-dependent translocation. Another inhibitor of the ribosomal translocation, thiostrepton, had no effect concerning the two activites, which is consistent with the fact that this antibiotic requires L11 for its binding to the ribosome. These results suggest that the ribosomes can perform a translocation reaction in the pyridine system, but in a factor-independent (spontaneous) manner.

ジャーナルBiological Chemistry
出版ステータスPublished - 1998 7月

ASJC Scopus subject areas

  • 生化学
  • 分子生物学
  • 臨床生化学


「A novel cell-free system for peptide synthesis driven by pyridine」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。