title: Protein-glycosphingolipid interactions revealed using catch-and-release mass spectrometry.
abstract: Glycosphingolipids (GSL) on the surface of cells are important receptors in antigen/microbial recognition and cell adhesion. However, their functional characterization is often challenging. We have developed a catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) assay for the identification of specific interactions between water-soluble proteins or protein complexes with GSL incorporated into nanodiscs. The specificity and sensitivity of the assay is demonstrated for interactions involving cholera toxin and Shiga toxin, with their natural GSL receptors, the ganglioside GM1, and the globotriaosylceramide Gb3, respectively. The detection of binding between cholera toxin and GM1 within a mixture of lipids extracted from cell membranes highlights the potential of this assay for the discovery of biologically relevant protein-GSL interactions.
authors.affiliation: Alberta Glycomics Centre, University of Alberta, Edmonton, Canada.
authors.forename: Yixuan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Lan
authors.initials: L
authors.lastname: Liu
authors.forename: Rambod
authors.initials: R
authors.lastname: Daneshfar
authors.forename: Elena N
authors.initials: EN
authors.lastname: Kitova
authors.forename: Caishun
authors.initials: C
authors.lastname: Li
authors.forename: Feng
authors.initials: F
authors.lastname: Jia
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Cairo
authors.forename: John S
authors.initials: JS
authors.lastname: Klassen
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D002772:Cholera Toxin / Q000378:metabolism*; D005677:G(M1) Ganglioside / Q000378:metabolism; D006028:Glycosphingolipids / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D022621:Shiga Toxin / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D014281:Trihexosylceramides / Q000378:metabolism
chemical_list: D006028:Glycosphingolipids; D014281:Trihexosylceramides; D005677:G(M1) Ganglioside; C018549:globotriaosylceramide; D022621:Shiga Toxin; D002772:Cholera Toxin