Alberta Glycomics Centre, University of Alberta, Edmonton, Canada.
Anal Chem. 2012 Sep 18;84(18):7618-21. doi: 10.1021/ac3023857. Epub 2012 Sep 4.
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.
细胞表面的糖脂（GSL）是抗原/微生物识别和细胞黏附中的重要受体。然而，其功能表征通常具有挑战性。我们开发了一种用于鉴定水溶性蛋白或蛋白复合物与纳米盘结合的 GSL 之间特定相互作用的捕获和释放电喷雾电离质谱（CaR-ESI-MS）测定法。该测定法的特异性和灵敏度已通过霍乱毒素和志贺毒素与其天然 GSL 受体神经节苷脂 GM1 和Globotriaosylceramide Gb3 之间的相互作用得到证明。在细胞膜提取的脂质混合物中检测到霍乱毒素与 GM1 之间的结合，突出了该测定法在发现具有生物学相关性的蛋白-GSL 相互作用方面的潜力。
