Abstract:
Protein N-glycosylation is an essential posttranslational modification which is initiated in the endoplasmic reticulum (ER). In plants, the N-glycans play a pivotal role in protein folding and quality control. Through the interaction of glycan processing and binding reactions mediated by ER-resident glycosidases and specific carbohydrate-binding proteins, the N-glycans contribute to the adoption of a native protein conformation. Properly folded glycoproteins are released from these processes and allowed to continue their transit to the Golgi where further processing and maturation of N-glycans leads to the formation of more complex structures with different functions. Incompletely folded glycoproteins are removed from the ER by a highly conserved degradation process to prevent the accumulation or secretion of misfolded proteins and maintain ER homeostasis. Here, we describe methods to analyze the N-glycosylation status and the glycan-dependent ER-associated degradation process in plants.
摘要:
蛋白质N-糖基化是在内质网(ER)中启动的必需的翻译后修饰。在植物中,N-聚糖在蛋白质折叠和质量控制中起关键作用。通过由ER驻留的糖苷酶和特定的碳水化合物结合蛋白介导的聚糖加工和结合反应的相互作用,N-聚糖有助于采用天然蛋白质构象。正确折叠的糖蛋白从这些过程中释放,并允许其继续转运到高尔基体,在那里N-聚糖的进一步加工和成熟导致形成具有不同功能的更复杂结构。不完全折叠的糖蛋白通过高度保守的降解过程从ER中去除,以防止错误折叠的蛋白质的积累或分泌并维持ER稳态。这里,我们描述了分析植物中N-糖基化状态和聚糖依赖性ER相关降解过程的方法。