Mucin-lectin interactions assessed by flow cytometry
Mucin-lectin interactions assessed by flow cytometry
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DOI：
10.1016/j.carres.2010.05.012
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发表时间：
2010-07-02
影响因子：
3.1
通讯作者：
Juge, Nathalie
中科院分区：
文献类型：
--
作者：
Jeffers, Faye;Fuell, Christine;Juge, Nathalie
粘蛋白和粘蛋白型糖蛋白的o糖基化结构域含有50-80%的碳水化合物，并具有扩展的构象。在这里，我们描述了一种流式细胞术（FCM）方法来确定凝集素与粘蛋白的碳水化合物结合特异性。将生物素化的粘蛋白固定在链霉亲和素包被的珠上，用GC-MS和MALDI-ToF测定主要粘蛋白糖链的结合特异性，并用荧光素标记的凝集素监测。作为对照，测定凝集素对特定生物素化聚糖的特异性。灵活，多参数数据采集，速度，灵敏度和高通量能力的优势使流式细胞术成为研究聚糖和蛋白质之间各种相互作用的有价值的工具。(c) 2010 Elsevier Ltd.版权所有。
The O-glycosylated domains of mucins and mucin-type glycoproteins contain 50-80% of carbohydrate and possess expanded conformations. Herein, we describe a flow cytometry (FCM) method for determining the carbohydrate-binding specificities of lectins to mucin. Biotinylated mucin was immobilized on streptavidin-coated beads, and the binding specificities of the major mucin sugar chains, as determined by GC-MS and MALDI-ToF, were monitored using fluorescein-labeled lectins. The specificities of lectins toward specific biotinylated glycans were determined as controls. The advantage of flexibility, multiparametric data acquisition, speed, sensitivity, and high-throughput capability makes flow cytometry a valuable tool to study diverse interactions between glycans and proteins. (c) 2010 Elsevier Ltd. All rights reserved.