Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322, USA.
Anal Biochem. 2012 Oct 1;429(1):82-7. doi: 10.1016/j.ab.2012.06.029. Epub 2012 Jul 6.
A major limitation in studying the structures and functions of glycans in glycosphingolipids is the difficulty in releasing free glycans for analysis and derivatization. Here we show that reducing glycans can be released nonenzymatically from glycosphingolipids after a brief treatment with ozone followed by heating in neutral aqueous buffer (pHs 6.0-8.0). The released free reducing glycans are then available for glycomic analyses, including fluorescent labeling, permethylation, and mass spectrometry. This procedure is simple and highly efficient, with no base-catalyzed "peeling" reaction by-products observed.
研究糖脂中的聚糖结构和功能的一个主要限制因素是难以释放游离聚糖进行分析和衍生化。在这里，我们表明，在臭氧短暂处理后，通过在中性水缓冲液（pH 值 6.0-8.0）中加热，可以非酶促方式从糖脂中释放出还原聚糖。然后，释放出的游离还原聚糖可用于糖组学分析，包括荧光标记、全甲基化和质谱分析。该方法简单高效，没有观察到碱催化的“剥皮”反应副产物。
