Gentamicin interaction with Pseudomonas aeruginosa cell envelope.

Gentamicin, an aminoglycoside antibiotic known to inhibit protein synthesis, had a detrimental effect on the integrity of the cell wall of Pseudomonas aeruginosa ATCC 9027 (a susceptible strain) as shown by electron microscopy using negative-staining, thin-sectioning, and freeze-fracture techniques. The disruption occurred in a sequential manner, moving from the outer membrane to the inner membrane, and could result in lysis of the cell. During this process the outer membrane lost 34% of its total protein and 30% of its lipopolysaccharide (measured as 2-keto-3-deoxyoctonate) upon exposure to 25 micrograms of gentamicin per ml for 15 min. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the outer membrane proteins showed altered banding patterns after exposure to gentamicin. Atomic absorption spectrophotometry revealed a decrease in magnesium and calcium content (18 and 38%, respectively) in the cell envelopes after gentamicin treatment. It is proposed that gentamicin displaces essential metal cations within the outer membrane, consequently destabilizing and extracting organic constituents. Small transient holes are thereby produced which make the outer membrane more permeable to the antibiotic and which expose the protoplast to high concentrations of gentamicin. This membrane effect may contribute to the effects of protein synthesis inhibition during the killing process.