RESUMO
The electrical potential difference (delta psi) across the membrane of Escherichia coli was measured by the distribution of lipid-soluble cations and correlated with resistance to dihydrostreptomycin, where resistance is presumed due to reduced uptake of the drug. A good correlation between the two measured parameters was found under all conditions tested, which included effects of several mutations, inhibitors, changes in pH, and osmolarity. The most dramatic changes were seen when pH was varied; in wild-type strains resistance increased more than 100-fold, and delta psi fell by 70 mV when pH was reduced from 8.5 to 5.5. These results were interpreted as support for a model in which the uptake of the polycationic aminoglycosides is electrogenic and therefore driven by delta psi. The factor common to mutations and conditions which increase resistance was a reduction in delta psi. A simple model was developed which relates the minimal inhibitory concentration to the rate of aminoglycoside uptake and the rate of growth.
Assuntos
Antibacterianos/farmacologia , Membrana Celular/fisiologia , Resistência Microbiana a Medicamentos , Escherichia coli/efeitos dos fármacos , 2,4-Dinitrofenol , Aminoglicosídeos/farmacologia , Sulfato de Di-Hidroestreptomicina/farmacologia , Dinitrofenóis/farmacologia , Escherichia coli/ultraestrutura , Potenciais da Membrana , Testes de Sensibilidade MicrobianaRESUMO
The accumulation of 204T1+ by Escherichia coli occurs primarily via either of two K+ transport systems called Kdp and TrkA. T1+ influx is inhibited and T1+ efflux is stimulated by the addition of K+ to the assay medium. Mutants defective in both the Kdp and TrkA systems accumulate little T1+. Uptake of triphenylmethylphosphonium, a lipid-soluble cation whose distribution is widely used to estimate the membrane electrical potential in bacteria, occurs to about the same extent in mutants that accumulate little T1+ as in strains that accumulate T1+ to high levels. These findings indicate that T1+ may be useful as a probe of bacterial K+ transport systems but is not a reliable indicator of the membrane electrical potential in E. coli.