Journal of Antimicrobial Chemotherapy, Vol 42, 597-603, Copyright © 1998 by The British Society for Antimicrobial Chemotherapy
KJ Williams and LJ Piddock
A centrifugation method was used to investigate the accumulation of 14C-
rifampicin by Staphylococcus aureus and Escherichia coli, and to
characterize the mechanism of rifampicin transport into S. aureus. For both
species, drug accumulation was rapid with the steady-state concentration
(SSC) reached within 40 s of drug exposure. Rifampicin accumulation by S.
aureus was temperature and pH dependent; the lower the experimental
temperature and the lower the experimental pH, the lower was the
concentration of rifampicin accumulated. Accumulation was unaffected by the
presence of inhibitors of antibiotic efflux, carbonyl cyanide
m-chlorophenylhydrazone (CCCP), dinitrophenol (DNP), or reserpine. Exposure
to increasing concentrations of rifampicin suggested that the accumulation
process was saturable above a rifampicin concentration of 0.2 mg/L.
Michaelis-Menten kinetics gave an apparent Km and Vmax for rifampicin,
calculated from a Lineweaver-Burk plot, of 0.05 mg/L (0.06 microM) and 3.8
ng rifampicin per second, respectively. However, calculations suggest that
these values reflect those for binding of rifampicin to its target, RNA
polymerase.
ORIGINAL ARTICLES
Accumulation of rifampicin by Escherichia coli and Staphylococcus aureus
Department of Infection, The Medical School, University of Birmingham, Edgbaston, UK.
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