使用体外甘氨基化在体外甘氨基化测试的细分标记糖蛋白的NMR结构确定
Vadim Slynko1, Mario Schubert, Shin Numao
1Institute of Molecular Biology and Biophysics, ETH Zurich, CH-8093 Zurich, Switzerland.
Journal of the American Chemical Society
|January 22, 2009
概括
这项研究提出了一种新的NMR方法,用于确定糖蛋白的3D结构. 该技术能够实现高度均的糖化,从而产生复杂的寡糖的明确结构.
科学领域:
- 生物化学 生物化学
- 结构生物学 结构生物学
- 葡萄糖科学 (Glycoscience) 是一种科学.
背景情况:
- 鉴定葡萄糖蛋白和复杂的寡糖的三维结构是具有挑战性的,因为问题,如不均的糖化和糟糕的结晶.
- 现有的方法与严重的化学转移重叠作斗争,限制结构分析.
研究的目的:
- 开发一种新的核磁共振 (NMR) 方法,用于结构性确定N-链 glycoproteins.
- 克服当前技术的局限性,使复杂的寡糖结构的分析.
主要方法:
- 通过体外糖化处理利用了甘氨酸和蛋白质成分的细分标记.
- 开发了一种基于NMR的新方法来确定糖蛋白的结构.
主要成果:
- 在毫克的蛋白质量中实现高度均的糖化.
- 成功确定了来自Campylobacter jejuni.的N链 glycoprotein 的结构.
- 在一个灵活的循环中确定了糖化受体位.
- 扩展了可观测的核过度干扰效应 (NOE) 距离限制,使寡糖化物超过4 Å,从而产生了每种糖化物链接的众多距离限制.
- 得到了一个明确的甘氨酸结构.
结论:
- 新的NMR方法显著改善了N链 glycoproteins的结构确定.
- 该技术可以更轻松地分析复杂的寡糖结构,克服以前的局限性.
- 这一进步提供了一种可靠的方法,用于获得高分辨率的结构,具有挑战性的糖蛋白标.
相关概念视频
01:24Oligosaccharide 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...
01:25Protein 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...