title: Electrochemical and PM-IRRAS characterization of cholera toxin binding at a model biological membrane.
abstract: A mixed phospholipid-cholestrol bilayer, with cholera toxin B (CTB) units attached to the monosialotetrahexosylganglioside (GM1) binding sites in the distal leaflet, was deposited on a Au(111) electrode surface. Polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) measurements were used to characterize structural and orientational changes in this model biological membrane upon binding CTB and the application of the electrode potential. The data presented in this article show that binding cholera toxin to the membrane leads to an overall increase in the tilt angle of the fatty acid chains; however, the conformation of the bilayer remains relatively constant as indicated by the small decrease in the total number of gauche conformers of acyl tails. In addition, the bound toxin caused a significant decrease in the hydration of the ester group contained within the lipid bilayer. Furthermore, changes in the applied potential had a minimal effect on the overall structure of the membrane. In contrast, our results showed significant voltage-dependent changes in the average orientation of the protein α-helices that may correspond to the voltage-gated opening and closing of the central pore that resides within the B subunit of cholera toxin.
authors.affiliation: Department of Chemistry, University of Guelph, Guelph, Ontario, Canada.
authors.forename: J Jay
authors.initials: JJ
authors.lastname: Leitch
authors.forename: Christa L
authors.initials: CL
authors.lastname: Brosseau
authors.forename: Sharon G
authors.initials: SG
authors.lastname: Roscoe
authors.forename: Kyrylo
authors.initials: K
authors.lastname: Bessonov
authors.forename: John R
authors.initials: JR
authors.lastname: Dutcher
authors.forename: Jacek
authors.initials: J
authors.lastname: Lipkowski
mesh_terms: D001665:Binding Sites; D002772:Cholera Toxin / Q000737:chemistry*; D055664:Electrochemical Techniques; D004566:Electrodes; D005732:Gangliosides / Q000737:chemistry*; D006046:Gold / Q000737:chemistry; D008958:Models, Molecular; D013055:Spectrophotometry, Infrared; D013499:Surface Properties
chemical_list: D005732:Gangliosides; D006046:Gold; D002772:Cholera Toxin