Greatly improved activity of staphylococcal ribosomes in polyadenylate directed polylysine synthesis: as an assay system for investigating their sensitivity to macrolide antibiotics.
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.
Nakajima
Y
Y
Division of Microbiology, Hokkaido Institute of Pharmaceutical Sciences, Japan.
Takeda
R
R
Tani
K
K
Endou
K
K
Matsuoka
M
M
Yamagishi
S
S
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D055231:Lincosamides; D018942:Macrolides; D008826:Microbial Sensitivity Tests / Q000379:methods; D010452:Peptide Biosynthesis; D010455:Peptides; D011061:Poly A / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism* / Q000502:physiology; D013211:Staphylococcus aureus / Q000378:metabolism / Q000648:ultrastructure*; D014769:Virginiamycin / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D055231:Lincosamides; D018942:Macrolides; D010455:Peptides; D014769:Virginiamycin; D011061:Poly A; C027819:polyleucine; C012001:polyphenylalanine