Minic Zoran, Jamet Elisabeth, Négroni Luc, Arsene der Garabedian P, Zivy Michel, Jouanin Lise
Laboratoire de Biologie Cellulaire, Institut National de la Recherche Agronomique, Route de Saint-Cyr, F-78026, Versailles Cedex, France.
J Exp Bot. 2007;58(10):2503-12. doi: 10.1093/jxb/erm082. Epub 2007 May 26.
N-glycosylated proteins were isolated from Arabidopsis thaliana mature stems using affinity chromatography on Concanavalin A Sepharose, separated by 2D-electrophoresis and identified using nanoHPLC-MS/MS and MALDI-TOF MS. 102 glycoproteins were identified. 94% of these proteins were predicted by bioinformatics to be targeted to the secretory pathway and 87% of them were predicted to be localized in the cell wall or at the plasma membrane. 30% of these proteins belong to glycoside hydrolase (GH) families with some of them possibly involved in the hydrolysis of cell wall polysaccharides. The second major class of identified proteins comprises aspartyl and serine proteases. Other proteins are predicted to be oxido-reductases, contain interacting domains, are potentially involved in signalling or have an unknown function. This is, to our knowledge, the first survey of plant cell wall N-glycosylated proteins.
使用伴刀豆球蛋白A琼脂糖亲和层析从拟南芥成熟茎中分离出N-糖基化蛋白,通过二维电泳进行分离,并使用纳升液相色谱-串联质谱(nanoHPLC-MS/MS)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)进行鉴定。共鉴定出102种糖蛋白。通过生物信息学预测,其中94%的蛋白靶向分泌途径,87%的蛋白预计定位于细胞壁或质膜。这些蛋白中有30%属于糖苷水解酶(GH)家族,其中一些可能参与细胞壁多糖的水解。鉴定出的第二大类蛋白包括天冬氨酸蛋白酶和丝氨酸蛋白酶。其他蛋白预计为氧化还原酶,含有相互作用结构域,可能参与信号传导或功能未知。据我们所知,这是对植物细胞壁N-糖基化蛋白的首次调查。