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INTERACTION OF AMINOGLYCOSIDES WITH THE OUTER MEMBRANES AND PURIFIED LIPOPOLYSACCHARIDE AND OMPF PORIN OF ESCHERICHIA-COLI
被引:127
作者:
HANCOCK, REW
FARMER, SW
LI, ZS
POOLE, K
机构:
[1] Department of Microbiology, University of British Columbia, Vancouver, BC
关键词:
D O I:
10.1128/AAC.35.7.1309
中图分类号:
Q93 [微生物学];
学科分类号:
071005 ;
100705 ;
摘要:
The mechanism of uptake of aminoglycosides across the outer membrane of Escherichia coli was reevaluated. Porin-deficient mutants showed no alteration in gentamicin or kanamycin susceptibility. Furthermore, the influence of kanamycin on intrinsic tryptophan fluorescence of porin OmpF (Y. Kobayashi, and T. Nakae, Eur. J. Biochem. 151:231-236, 1985) was shown to be strongly influenced by protein concentration and EDTA. This led to the hypothesis that aminoglycoside-mediated increases and decreases in intrinsic tryptophan fluorescence were due to aggregation-disaggregation of OmpF mediated by interaction at a divalent cation binding site on OmpF. Gentamicin, kanamycin, and polymyxin B increased E. coli outer membrane permeability to the hydrophobic fluorescent compound 1-N-phenyl-naphthylamine (NPN) and the peptidoglycan-degrading enzyme lysozyme. Addition of Mg2+ blocked these permeabilizing activities. Furthermore, gentamicin and polymyxin B bound to Mg2+-binding sites on E. coli lipopolysaccharide, as determined in dansyl polymyxin displacement experiments. A polymyxin-resistant, lipopolysaccharide-altered pmr mutant of E. coli had a fourfold-lower MIC of gentamicin and kanamycin and was more poorly permeabilized to 1-N-phenyl-naphthylamine than was its parent strain. These data were consistent with uptake of aminoglycosides across the E. coli outer membrane by the self-promoted uptake mechanism.
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页码:1309 / 1314
页数:6
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