The bactericidal action of streptomycin: membrane permeabilization caused by the insertion of mistranslated proteins into the cytoplasmic membrane of Escherichia coli and subsequent caging of the antibiotic inside the cells due to degradation of these proteins.
The mechanism by which the aminoglycoside antibiotic streptomycin permeabilizes the cytoplasmic membrane of Escherichia coli cells was reinvestigated. For this purpose, the extent of streptomycin-induced K+ loss from cells growing at low external K+ concentrations was taken as a measure of membrane permeabilization. Experiments with different K(+)-uptake mutants showed that the antibiotic specifically increased the passive permeability of the cell membrane to K+ and other ions. These permeability changes were small and the membrane potential of the treated cells remained high. The membrane permeabilization was not due to a direct interaction of the antibiotic with the cell membrane, since cells that carry an rpsL mutation and synthesize proteins in a streptomycin-insensitive way did not lose K+ after the addition of the antibiotic. Due to misreading and premature termination of translation the cells synthesized aberrant proteins under the conditions where membrane permeabilization occurred. Two conditions are described under which the cells both degraded these mistranslated proteins rapidly and reaccumulated K+, lending support to the hypothesis that membrane permeabilization is due to the presence of the mistranslated proteins in the cell membrane. Evidence is presented that the irreversibility of (dihydro)streptomycin uptake by cells washed free from the antibiotic might also be due to rapid degradation of the mistranslated proteins, leading to 'caging' of the antibiotic inside the cells.
Busse
H J
HJ
Universität Osnabrück, Fachgebiet Mikrobiologie, FRG.
Wöstmann
C
C
Bakker
E P
EP
D001692:Biological Transport; D002455:Cell Division / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects*; D004096:Dihydrostreptomycin Sulfate / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects* / Q000378:metabolism; D029968:Escherichia coli Proteins; D008024:Ligands; D008564:Membrane Potentials; D009154:Mutation; D011188:Potassium / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D000091243:Ribosomal Protein S9; D012413:Rubidium / Q000378:metabolism; D013307:Streptomycin / Q000494:pharmacology*
D029968:Escherichia coli Proteins; D008024:Ligands; D011500:Protein Synthesis Inhibitors; D000091243:Ribosomal Protein S9; C000717190:RpsI protein, E coli; D012413:Rubidium; D011188:Potassium; D004096:Dihydrostreptomycin Sulfate; D013307:Streptomycin