Toxin studies using an integrated biophysical and structural biology approach.

Last, Julie A. ; Schroeder, Anne E. ; Slade, Andrea Lynn ; Sasaki, Darryl Yoshio ; Yip, Christopher M. (University of Toronto, Toronto, Ontario, Canada) ; Schoeniger, Joseph S. (Sandia National Laboratories, Livermore, CA)

Sandia National Laboratories

关键词: Membranes;    Toxins Molecular Biology-Methodology.;    Biology.;    Gangliosides;    Tetanus;   

DOI  :  10.2172/922768
RP-ID  :  SAND2005-1700
RP-ID  :  AC04-94AL85000
RP-ID  :  922768

美国|英语

来源: UNT Digital Library

【 摘 要 】

Clostridial neurotoxins, such as botulinum and tetanus, are generally thought to invade neural cells through a process of high affinity binding mediated by gangliosides, internalization via endosome formation, and subsequent membrane penetration of the catalytic domain activated by a pH drop in the endosome. This surface recognition and internalization process is still not well understood with regard to what specific membrane features the toxins target, the intermolecular interactions between bound toxins, and the molecular conformational changes that occur as a result of pH lowering. In an effort to elucidate the mechanism of tetanus toxin binding and permeation through the membrane a simple yet representative model was developed that consisted of the ganglioside G{sub tlb} incorporated in a bilayer of cholesterol and DPPC (dipalmitoylphosphatidyl choline). The bilayers were stable over time yet sensitive towards the binding and activity of whole toxin. A liposome leakage study at constant pH as well as with a pH gradient, to mimic the processes of the endosome, was used to elucidate the effect of pH on the toxin's membrane binding and permeation capability. Topographic imaging of the membrane surface, via in situ tapping mode AFM, provided nanoscale characterization of the toxin's binding location and pore formation activity.