PMID:1656859

Citation	Hancock, RE, Farmer, SW, Li, ZS and Poole, K (1991) Interaction of aminoglycosides with the outer membranes and purified lipopolysaccharide and OmpF porin of Escherichia coli.Antimicrob. Agents Chemother. 35:1309-14
Abstract	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 Mg(2+)-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-phenylnaphthylamine 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.
Links	PubMed
Keywords	Anti-Bacterial Agents; Bacterial Outer Membrane Proteins; Culture Media; Escherichia coli; Gentamicins; Kanamycin; Lipopolysaccharides; Polymyxin B; Porins
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Phenotype of	Taxon Information	Genotype Information (if known)	Condition Information	OMP ID	OMP Term Name	ECO ID	ECO Term Name	Notes	Status
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SC9252 NCBI Taxon ID: 83333	Genotype of Reference Strain: pmrA Genotype of Experimental Strain : pmrA	Reference Condition:	OMP:0000276	increased resistance to antimicrobial compound	ECO:0000182	in vitro culture assay data	Resistant to polymyxin, table 1
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SC9252 NCBI Taxon ID: 83333	Genotype of Reference Strain: pmrA Genotype of Experimental Strain : pmrA	Reference Condition:	OMP:0000276	increased resistance to antimicrobial compound	ECO:0000182	in vitro culture assay data	Resistant to Gentamicin, table 1
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References

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