How beta-lactam antibiotics enter bacteria: a dialogue with the porins.
BACKGROUND
Multi-drug resistant (MDR) infections have become a major concern in hospitals worldwide. This study investigates membrane translocation, which is the first step required for drug action on internal bacterial targets. beta-lactams, a major antibiotic class, use porins to pass through the outer membrane barrier of Gram-negative bacteria. Clinical reports have linked the MDR phenotype to altered membrane permeability including porin modification and efflux pump expression.


RESULTS
Here influx of beta-lactams through the major Enterobacter aerogenes porin Omp36 is characterized. Conductance measurements through a single Omp36 trimer reconstituted into a planar lipid bilayer allowed us to count the passage of single beta-lactam molecules. Statistical analysis of each transport event yielded the kinetic parameters of antibiotic travel through Omp36 and distinguishable translocation properties of beta-lactams were quantified for ertapenem and cefepime. Expression of Omp36 in an otherwise porin-null bacterial strain is shown to confer increases in the killing rate of these antibiotics and in the corresponding bacterial susceptibility.


CONCLUSIONS
We propose the idea of a molecular "passport" that allows rapid transport of substrates through porins. Deciphering antibiotic translocation provides new insights for the design of novel drugs that may be highly effective at passing through the porin constriction zone. Such data may hold the key for the next generation of antibiotics capable of rapid intracellular accumulation to circumvent the further development MDR infections.
James
Chloë E
CE
UMR-MD-1, Transporteurs membranaires, Chimiorésistance et Drug Design, Faculté de Médecine, IFR 88, Université de la Méditerranée, Marseille, France.
Mahendran
Kozhinjampara R
KR
Molitor
Alexander
A
Bolla
Jean-Michel
JM
Bessonov
Andrey N
AN
Winterhalter
Mathias
M
Pagès
Jean-Marie
JM
D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism; D007700:Kinetics; D008051:Lipid Bilayers / Q000378:metabolism; D018272:Porins / Q000378:metabolism*; D047090:beta-Lactams / Q000378:metabolism* / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D008051:Lipid Bilayers; D018272:Porins; D047090:beta-Lactams