Effects of glycosylation on protein structure, function and dynamics

糖基化对蛋白质结构、功能和动力学的影响

• 批准号: 347211955
• 负责人: Professor Dr. Harald Schwalbe
• 金额: --
• 依托单位: Institut für Organische Chemie und Chemische Biologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2023-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/347211955?language=en
• 关键词: Effects; glycosylation; protein; structure; function

Recent years have seen remarkable progress in the field of structural biology of proteins. However, human or complex proteins with posttranslational modifications (like glycosylation, phosphorylation and methylation etc.) are rarely found in the Protein Data Bank (PDB), since these modifications cannot be performed sufficiently by prokaryotic protein expression hosts like Escherichia coli. Among the posttranslational modifications, protein glycosylation is the most abundant protein modification found in nature, introducing more diversity into the protein than all the other posttranslational modifications combined. Glycosylation of proteins changes their structure, thermodynamics and kinetics, which modulate the function of the proteins and influence their localization, trafficking, solubility, antigenicity, biological activity, half-life and cell-cell interactions. Therefore, it is not surprising that glycan structures and glycosylation patterns are highly dynamic and change during development and in certain diseases. So far, the effects of protein glycosylation on function are poorly understood. Even more, structural information regarding this posttranslational modification is rare. In fact, from a structural biology perspective glycosylation is undesired and is often considered as an additional obstacle to solve a protein structure due to the size, flexibility and heterogeneity of the oligosaccharide chains. In the Research Unit our group (P6 Schwalbe) wants to study the influence of glycosylation on the structure, function and dynamics of glycoproteins by NMR spectroscopy. For this purpose, we will develop specific NMR tools (expression, isotope labeling, NMR methods) to conduct research on glycosylated proteins in the first funding period. In the Research Unit we will focus on three highly conserved glycosylation pathways (N-glycosylation, C-mannosylation and O-mannosylation) which are based on the lipid dolichol and compete for both mannosyl donor substrates and acceptor proteins. In the course of the assembly of this Research Unit we already started to structurally investigate the effect of tryptophan C-mannosylation by NMR in collaboration with (P1 Bakker).

近年来，蛋白质结构生物学领域取得了显著进展。然而，具有翻译后修饰（如糖基化、磷酸化和甲基化等）的人类或复杂蛋白质，在蛋白质数据库（PDB）中很少发现，因为这些修饰不能通过原核蛋白质表达宿主如大肠杆菌充分进行。在翻译后修饰中，蛋白质糖基化是自然界中发现的最丰富的蛋白质修饰，它比所有其他翻译后修饰的组合为蛋白质引入了更多的多样性。蛋白质的糖基化改变了蛋白质的结构、热力学和动力学，从而调节蛋白质的功能，并影响蛋白质的定位、运输、溶解性、抗原性、生物活性、半衰期和细胞-细胞相互作用。因此，聚糖结构和糖基化模式在发育过程中和某些疾病中是高度动态和变化的，这并不奇怪。到目前为止，人们对蛋白质糖基化对功能的影响知之甚少。更重要的是，关于这种翻译后修饰的结构信息是罕见的。事实上，从结构生物学的角度来看，糖基化是不期望的，并且由于寡糖链的大小、柔性和异质性，通常被认为是解决蛋白质结构的额外障碍。在研究单元中，我们的小组（P6 Schwalbe）希望通过NMR光谱研究糖基化对糖蛋白的结构，功能和动力学的影响。为此，我们将开发特定的NMR工具（表达，同位素标记，NMR方法），以在第一个资助期内对糖基化蛋白质进行研究。在研究单元中，我们将专注于三个高度保守的糖基化途径（N-糖基化，C-甘露糖基化和O-甘露糖基化），这些途径基于脂质多萜醇并竞争甘露糖基供体底物和受体蛋白。在该研究单元的组装过程中，我们已经开始与（P1 Bakker）合作，通过NMR对色氨酸C-甘露糖基化的影响进行结构研究。