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Antimicrobial Agents and Chemotherapy, November 2000, p. 2969-2978, Vol. 44, No. 11
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Stages of Polymyxin B Interaction with the
Escherichia coli Cell Envelope
Rimantas
Daugelavi
ius,1,2,*
Elena
Bakien
,1 and
Dennis H.
Bamford2
Department of Biochemistry and Biophysics,
Vilnius University,
iurlionio 21, LT-2009 Vilnius,
Lithuania,1 and Institute of
Biotechnology and Department of Biosciences, FIN-00014 University
of Helsinki, Helsinki, Finland2
Received 2 May 2000/Returned for modification 3 June 2000/Accepted 31 July 2000
The effects of polymyxin B (PMB) on the Escherichia
coli outer (OM) and cytoplasmic membrane (CM) permeabilities
were studied by monitoring the fluxes of tetraphenylphosphonium,
phenyldicarbaundecaborane, and K+ and H+
ions. At concentrations between 2 and 20 µg/ml, PMB increased the OM
permeability to lipophilic compounds and induced a
leakage of K+ from the cytosol and an accumulation of
lipophilic anions in the cellular membranes but did not
cause the depolarization of the CM. At higher concentrations, PMB
depolarized the CM, forming ion-permeable pores in the cell envelope.
The permeability characteristics of PMB-induced pores mimic those of
bacteriophage- and/or bacteriocin-induced channels. However, the
bactericidal effect of PMB took place at concentrations below 20 µg/ml, indicating that this effect is not caused by pore formation.
Under conditions of increased ionic strength, PMB made the OM permeable
to lipophilic compounds and decreased the K+
gradient but was not able to depolarize the cells. The
OM-permeabilizing effect of PMB can be diminished by increasing the
concentration of Mg2+. The major new findings of this work
are as follows: (i) the OM-permeabilizing action of PMB was dissected
from its depolarizing effect on the CM, (ii) the PMB-induced
ion-permeable pores in bacterial envelope were registered, and (iii)
the pore formation and depolarization of the CM are not obligatory for
the bactericidal action of PMB and dissipation of the K+
gradient on the CM.
*
Corresponding author. Mailing address: Department
of Biosciences, Biocentre 2, P.O. Box 56 (Viikinkaari 5),
FIN-00014 University of Helsinki, Helsinki, Finland. Phone: 358 9 19159097. Fax: 358 9 19159098. E-mail:
daugelav{at}cc.helsinki.fi.
Antimicrobial Agents and Chemotherapy, November 2000, p. 2969-2978, Vol. 44, No. 11
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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