SUPRAMOLECULAR ASSEMBLIES |
In Situ Scanning Probe Microscopy Studies of
Tetanus Toxin-Membrane Interactions
Andrea Slade 1, Joseph Schoeniger 2, Darryl Sasaki 2 and Christopher M Yip 3*
1 Digital Instruments / Veeco
2 Sandia National Laboratories
3 University of Toronto
* To whom correspondence should be addressed. E-mail: christopher.yip{at}utoronto.ca.
Submitted on December 27, 2005
Revised on February 24, 2006
Accepted on 22 August 2006
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Abstract |
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Despite the considerable information available with regards to the structure of the clostridial neurotoxins, and their inherent threat as biological warfare agents, the mechanisms underpinning their interactions with and translocation through the cell membrane remain poorly understood. We report herein the results of an in situ scanning probe microscopy (SPM) study of the interaction of tetanus toxin C-fragment (Tet C) with supported planar lipid bilayers (SPBs) containing the ganglioside receptor GT1b. Our results show that Tet C preferentially binds to the surface of fluid phase domains within biphasic membranes containing GT1b and that with an extended incubation period these interactions lead to dramatic changes in the morphology of the lipid bilayer, including the formation of 40 - 80 nm diameter circular cavities. Combined AFM/TIRFM experiments confirmed the presence of Tet C in the membrane after extended incubation. These morphological changes were found to be dependent upon the presence of GT1b and the solution pH.
Key Words:
AFM, TIRF, ganglioside receptor, tetanus, toxin binding
