A new mechanism of antibiotic resistance in Enterobacteriaceae induced by a structural modification of the major porin.
In Enterobacter aerogenes, multidrug resistance involves a decrease in outer membrane permeability associated with changes in an as yet uncharacterized porin. We purified the major porin from the wild-type strain and a resistant strain. We characterized this porin, which was found to be an OmpC/OmpF-like protein and analysed its pore-forming properties in lipid bilayers. The porin from the resistant strain was compared with the wild-type protein and we observed (i) that its single-channel conductance was 70% lower than that of the wild type; (ii) that it was three times more selective for cations; (iii) a lack of voltage sensitivity. These results indicate that the clinical strain is able to synthesize a modified porin that decreases the permeability of the outer membrane. Mass spectrometry experiments identified a G to D mutation in the putative loop 3 of the porin. Given the known importance of this loop in determining the pore properties of porins, we suggest that this mutation is responsible for the novel resistance mechanism developed by this clinical strain, with changes in porin channel function acting as a new bacterial strategy for controlling beta-lactam diffusion via porins.
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UMR 6522, CNRS, IFRMP 23, Faculté des Sciences, 76821 Mont-Saint-Aignan Cedex, France.
Baslé
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A
Jaquinod
M
M
Saint
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N
Malléa
M
M
Molle
G
G
Pagès
J M
JM
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D002463:Cell Membrane Permeability; D018550:Cephalosporin Resistance; D002511:Cephalosporins / Q000494:pharmacology; D021902:Enterobacter aerogenes / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007473:Ion Channels / Q000502:physiology; D013058:Mass Spectrometry / Q000379:methods; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009154:Mutation; D018272:Porins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013329:Structure-Activity Relationship
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D002511:Cephalosporins; D007473:Ion Channels; D008565:Membrane Proteins; D018272:Porins