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Glycomic approaches to study post-translational modification by O-linked beta-N-acetylglucosamine (O-GlcNAc).

机译:糖基方法研究O联β-N-乙酰氨基葡萄糖(O-GlcNAc)的翻译后修饰。

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摘要

The overall goal of my research projects is to understand the complex interplay between post-translational O-GlcNAc and O-Phosphate modifications. Like phosphorylation, O-GlcNAcylation is a one of the major forms of post-translational modification (PTM), and it has been found on numerous nuclear and cytoplasmic proteins that are involved in diverse cellular metabolism and function. Similar to phosphorylation, O-GlcNAcylation occurs on serine or threonine residues of the polypeptide backbones. O-GlcNAcylation is dynamic and has a high cycling rate on many proteins in response to various environmental stimuli. So far, many proteins have been found to be both O-GlcNAcylated and phosphorylated. The "yin-yang" hypothesis states that competition of O-GlcNAc and O-phosphate on the same serine or threonine residue may modulate the function of the protein. However, accumulating evidence has suggested this hypothesis as over-simplified. Global and systematic studies have been proposed and performed using mass spectrometry-based proteomics approaches in order to elucidate the complex interplay between the two abundant post translational modifications. An important aspect of studying post-translational modifications is to map the exact sites of the modifications. Mapping O-GlcNAc sites has been extremely difficult due to their intrinsic lability in the gas phase, low stoichiometry, and signal suppression. To solve this problem, new methods for O-GlcNAc enrichment and site-mapping are developed and optimized.
机译:我的研究项目的总体目标是了解翻译后O-GlcNAc与O-磷酸盐修饰之间的复杂相互作用。像磷酸化一样,O-GlcNAcylation是翻译后修饰(PTM)的主要形式之一,并且已在涉及多种细胞代谢和功能的众多核蛋白和胞质蛋白上发现。与磷酸化相似,O-GlcNAcylation发生在多肽主链的丝氨酸或苏氨酸残基上。 O-GlcNAcylation是动态的,对多种蛋白质具有较高的循环速率,以响应各种环境刺激。迄今为止,已经发现许多蛋白质都被O-GlcNA酰化和磷酸化。 “阴阳”假说指出,O-GlcNAc和O-磷酸盐在同一丝氨酸或苏氨酸残基上的竞争可能会调节该蛋白的功能。但是,越来越多的证据表明这一假设过于简单。为了阐明两种丰富的翻译后修饰之间的复杂相互作用,已经提出并使用基于质谱的蛋白质组学方法进行了全球和系统的研究。研究翻译后修饰的一个重要方面是绘制修饰的确切位点。由于O-GlcNAc位点在气相中固有的不稳定性,低化学计量和信号抑制,因此对其进行定位非常困难。为了解决这个问题,开发并优化了O-GlcNAc富集和位点映射的新方法。

著录项

  • 作者

    Wang, Zihao.;

  • 作者单位

    The Johns Hopkins University.;

  • 授予单位 The Johns Hopkins University.;
  • 学科 Biology Molecular.;Chemistry Biochemistry.
  • 学位 Ph.D.
  • 年度 2009
  • 页码 207 p.
  • 总页数 207
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;生物化学;
  • 关键词

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