Protein鈥揋lycosphingolipid Interactions Revealed Using Catch-and-Release Mass Spectrometry

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• 作者：Yixuan Zhang ; Lan Liu ; Rambod Daneshfar ; Elena N. Kitova ; Caishun Li ; Feng Jia ; Christopher W. Cairo ; John S. Klassen
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：September 18, 2012
• 年：2012
• 卷：84
• 期：18
• 页码：7618-7621
• 全文大小：251K
• 年卷期：v.84,no.18(September 18, 2012)
• ISSN：1520-6882

文摘

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鈥揋SL interactions.