Molecular mechanisms of interaction of rabbit CAP18 with outer membranes of gram-negative bacteria.
The mechanism of interaction of the cationic antimicrobial protein (18 kDa), CAP18, with the outer membrane of Gram-negative bacteria was investigated applying transmission electron microscopy and voltage-clamp techniques on artificial planar bilayer membranes. Electron micrographs of bacterial cells exposed to CAP18 showed damage to the outer membrane of the sensitive Escherichia coli strains F515 and ATCC 11775, whereas the membrane of the resistant Proteus mirabilis strain R45 remained intact. Electrical measurements on various planar asymmetric bilayer membranes, one side consisting of a phospholipid mixture and the other of different phospholipids or of lipopolysaccharide (reconstitution model of the outer membrane), yielded information about the influence of CAP18 on membrane integrity. Addition of CAP18 to the side with the varying lipid composition led to lipid-specific adsorption of CAP18 and subsequent induction of current fluctuations due to the formation of transient membrane lesions at a lipid-specific clamp voltage. We propose that the applied clamp voltage leads to reorientation of CAP18 molecules adsorbed to the bilayer into an active transmembrane configuration, allowing the formation of lesions by multimeric clustering.
Gutsmann
T
T
Department of Immunochemistry and Biochemical Microbiology, Center for Medicine and Biosciences, Research Center Borstel, Germany.
Larrick
J W
JW
Seydel
U
U
Wiese
A
A
D000818:Animals; D000900:Anti-Bacterial Agents / Q000037:antagonists & inhibitors / Q000737:chemistry; D023181:Antimicrobial Cationic Peptides; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000502:physiology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000502:physiology; D054804:Cathelicidins; D004553:Electric Conductivity; D004926:Escherichia coli / Q000737:chemistry / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D006493:Heparin / Q000494:pharmacology; D008051:Lipid Bilayers / Q000737:chemistry; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors / Q000737:chemistry*; D008564:Membrane Potentials; D011513:Proteus mirabilis / Q000737:chemistry / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D011817:Rabbits
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D054804:Cathelicidins; D008051:Lipid Bilayers; D008070:Lipopolysaccharides; D006493:Heparin