从细胞溶解物中并行识别O-GlcNAc修饰的蛋白质
Hwan-Ching Tai1, Nelly Khidekel, Scott B Ficarro
1Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
Journal of the American Chemical Society
|August 26, 2004
概括
研究人员开发了一种新方法来识别O-GlcNAc-glycosylated蛋白质,从而能够快速分析任何细胞类型中的特定蛋白质和糖化位点. 这推动了对这一关键的翻译后修改的研究.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 蛋白质组学是指蛋白质组学.
背景情况:
- O-GlcNAcylation是一种动态的翻译后修饰,对细胞信号传递和调节至关重要.
- 鉴定O-GlcNAc-glycosylated蛋白质是具有挑战性的,因为其稀少性和动态性质.
- 现有的方法缺乏探测单个蛋白质的特异性和吞吐量.
研究的目的:
- 开发一种新的并行策略来识别O-GlcNAc-glycosylated蛋白质.
- 为了在各种细胞类型中快速检测O-GlcNAc修饰的特定蛋白质.
- 为了方便在上绘制糖化位的地图.
主要方法:
- 从细胞溶解物中并行识别O-GlcNAc-糖化蛋白的新策略.
- 该方法允许对特定感兴趣的蛋白质进行有针对性的分析.
- 该方法可以扩展到用于网站映射的.
主要成果:
- 成功识别了四种新的低丰度O-GlcNAc-糖化蛋白:c-Fos,c-Jun,ATF-1和CBP.
- 在O-GlcNAc转移酶 (OGT) 酶内确定了两个短糖化区域.
- 证明了在各种组织或细胞类型中准特定蛋白质的能力.
结论:
- 新的策略为研究O-GlcNAc糖化提供了一个强大的工具.
- 补充了现有的蛋白质组技术,用于蛋白质修饰分析.
- 预计将大大提高对O-GlcNAc在细胞过程中的作用的理解.
相关概念视频
Protein Glycosylation
Glycosylation, the most common post-translational modification for proteins, serves diverse functions. Adding sugars to proteins makes the proteins more resistant to proteolytic digestion. Glycosylated proteins can act as markers and receptors to promote cell-cell adhesion. Additionally, they have many essential quality control functions in the cell, such as correct protein folding and facilitating transport of misfolded proteins to the cytosol, which can be degraded.
Glycosylation occurs in...
Glycosylation occurs in...
Oligosaccharide Assembly
Protein glycosylation starts in the ER lumen and continues in the Golgi apparatus. Glycosyltransferases catalyze the addition of sugar molecules or glycosylation of proteins. Usually, these enzymes add sugars to the hydroxyl groups of selected serine or threonine residues to form O-linked glycans or the amino groups of asparagine residues to form N-linked glycans. Different positions on the same polypeptide chain can contain differently linked glycans.
Multiple sugar molecules that may or may...
Multiple sugar molecules that may or may...
Protein Folding Quality Check in the RER
ER is the primary site for the maturation and folding of soluble and transmembrane secretory proteins. The calnexin cycle is a specific chaperone system that folds and assesses the confirmation of N-glycosylated proteins before they can exit the ER lumen. The primary players of this quality check pipeline are the lectins, ER-resident chaperones, and a glucosyl transferase enzyme. In case the calnexin system in the lumen fails to salvage a misfolded protein, it is transported to the cytoplasm...
Proteoglycans
Glycans, a class of complex heterogeneous molecules, can be covalently attached to proteins to form glycosylated proteins that regulate various physiological and pathological processes. Glycosylated proteins or glycoproteins comprise N-linked and O-linked oligosaccharides. O-glycosylation is the most common type of protein glycosylation. Here, glycans attach to the oxygen atom of the hydroxyl groups of Serine or Threonine residues. O-linked glycosylation occurs later in protein processing,...


