title: Part I: an x-ray scattering study of cholera toxin penetration and induced phase transformations in lipid membranes.
abstract: Cholera toxin is a highly efficient biotoxin, which is frequently used as a tool to investigate protein-membrane interactions and as a reporter for membrane rafts. Cholera toxin binds selectively to gangliosides with highest affinity to GM(1). However, the mechanism by which cholera toxin crosses the membrane remains unresolved. Using x-ray reflectivity and grazing incidence diffraction, we have been able to monitor the binding and penetration of cholera toxin into a model lipid monolayer containing the receptor GM(1) at the air-water interface. Very high toxin coverage was obtained allowing precise measurements of how toxin binding alters lipid packing. Grazing incidence x-ray diffraction revealed the coexistence of two monolayer phases after toxin binding. The first was identical to the monolayer before toxin binding. In regions where toxin was bound, a second membrane phase exhibited a decrease in order as evidenced by a larger area per molecule and tilt angle with concomitant thinning of the monolayer. These results demonstrate that cholera toxin binding induces the formation of structurally distinct, less ordered domains in gel phases. Furthermore, the largest decrease in lateral order to the monolayer occurred at low pH, supporting a low endosomal pH in the infection pathway. Surprisingly, at pH = 8 toxin penetration by the binding portion of the toxin, the B(5) pentamer, was also observed.
authors: Manuel Lujan Neutron Scattering Center, Los Alamos National Laboratory, Los Alamos, New Mexico, USA.
authors: C E
authors: CE
authors: Miller
authors: J
authors: J
authors: Majewski
authors: E B
authors: EB
authors: Watkins
authors: T L
authors: TL
authors: Kuhl
mesh_terms: D002772:Cholera Toxin / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D008560:Membrane Fluidity; D008968:Molecular Conformation; D044367:Phase Transition; D014961:X-Ray Diffraction
chemical_list: D008051:Lipid Bilayers; D002772:Cholera Toxin