Greatly improved activity of staphylococcal ribosomes in polyadenylate directed polylysine synthesis: as an assay system for investigating their sensitivity to macrolide antibiotics

Journal of Pharmacobio-dynamics
Y NakajimaS Yamagishi

Abstract

In polyadenylate directed polylysine synthesis, homologously cell-free extracts containing ribosomes and S-100 (105,000 x g supernatant) from staphylococcal cells have less than one-half (one-tenth, when the extracts were stored at -80 degrees C within a few weeks) of the activity of the extracts from Escherichia coli Q13. The present study is concerned with further improving the activity of staphylococcal ribosomes. The polylysine-synthesizing ability by staphylococcal ribosomes increased up to about two times as much as that by E. coli Q13 ribosomes, when S-100 from E. coli Q13 was mixed with staphylococcal ribosomes which had been washed with a high salt HEPES buffer containing 10 mM HEPES, 1 mM EGTA, 16 mM magnesium acetate, 1.0 M ammonium chloride and 0.1 mM dithiothreitol (pH7.6). Polylysine synthesis by the heterologous extracts has an advantage over polyuridylate-directed polyphenylalanine synthesis in the analysis of ribosome sensitivity for macrolide antibiotics, especially erythromycin.

Citations

Jan 26, 2002·Journal of Infection and Chemotherapy : Official Journal of the Japan Society of Chemotherapy·Yoshinori Nakajima

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