Fitchette Anne-Catherine, Dinh Olivia Tran, Faye Loïc, Bardor Muriel
CNRS UMR 6037, IFRMP 23, GDR 2590, Université de Rouen, Mont Saint-Aignan, France.
Methods Mol Biol. 2007;355:317-42. doi: 10.1385/1-59745-227-0:317.
In plant cells, as in other eucaryotic cells, glycosylation is one of the most studied posttranslational events. It can be of two types, N- or O-glycosylation, depending on the linkage involved between the protein backbone and the oligosaccharide moiety. In this review, we present different methods, commonly used in our laboratory, to study the glycosylation of plant proteins. These approaches rely on blot detection with glycan-specific probes, as well as specific deglycosylation of the glycoproteins, followed by mass spectrometry analysis. Such experiments not only allow determination of whether the protein is a glycoprotein, but also how and where it is glycosylated. The last part of this chapter is dedicated to the specific purification and identification of glycoprotein populations in plant cells, so-called glycoproteomics.
与其他真核细胞一样，在植物细胞中，糖基化是研究最多的翻译后事件之一。根据蛋白质主链与寡糖部分之间的连接方式，糖基化可分为两种类型，即N - 糖基化或O - 糖基化。在本综述中，我们介绍了我们实验室常用的不同方法，用于研究植物蛋白的糖基化。这些方法依赖于用聚糖特异性探针进行印迹检测，以及对糖蛋白进行特异性去糖基化，随后进行质谱分析。此类实验不仅可以确定该蛋白是否为糖蛋白，还能确定其糖基化的方式和位置。本章的最后一部分专门介绍植物细胞中糖蛋白群体的特异性纯化和鉴定，即所谓的糖蛋白质组学。
