Interaction of cytolytic toxin CytB with a supported lipid bilayer: study using an acoustic wave device.
An acoustic technique was used to monitor the interaction of the pore-forming cytolytic toxin CytB with a positively charged supported lipid bilayer. The acoustic device, which is based on a waveguide geometry, is sensitive to changes in the mass of the supported bilayer. The specificity of the interaction, rate and extent of the association, reversibility and effect of previous depositions of toxin were investigated. The CytB was found to bind irreversibly to the lipids at all fractional coverages even when the protein-to-lipid ratio was high enough to imply that the protein was associating with the external surface of the bilayer. The CytB formed stable structures with the bilayer at high protein surface concentrations and did not appear to disrupt the bilayer in the manner of a detergent. The rate of association with the bilayer was found to be directly proportional to the solution concentration of CytB at higher concentrations but appeared to be low at a CytB solution concentration of 5 microg mL(-1), leading to relatively low amounts of CytB being associated with the bilayer.
Melzak
Kathryn A
KA
Institute of Biotechnology, University of Cambridge, Cambridge, United Kingdom.
Ellar
David J
DJ
Gizeli
Electra
E
D000162:Acoustics; D001413:Bacillus thuringiensis / Q000378:metabolism; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry*; D001427:Bacterial Toxins / Q000737:chemistry*; D001703:Biophysics / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D004731:Endotoxins / Q000737:chemistry*; D006460:Hemolysin Proteins; D007700:Kinetics; D008051:Lipid Bilayers / Q000378:metabolism*; D008055:Lipids / Q000737:chemistry; D015233:Models, Statistical; D019904:Polymethyl Methacrylate / Q000737:chemistry; D011485:Protein Binding; D013016:Sound; D013499:Surface Properties; D013997:Time Factors; D014118:Toxins, Biological / Q000737:chemistry
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D008051:Lipid Bilayers; D008055:Lipids; D014118:Toxins, Biological; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D019904:Polymethyl Methacrylate