Quantitative analysis of O-GlcNAcylation in combination with isobaric tag labeling and chemoenzymatic enrichment
Quantitative analysis of O-GlcNAcylation in combination with isobaric tag labeling and chemoenzymatic enrichment
复制标题
结合同量异序标签标记和化学酶富集对 O-GlcNAc 酰化进行定量分析
DOI：
10.1016/j.bmcl.2017.10.005
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发表时间：
2017
影响因子：
2.7
通讯作者：
Yuri Miura.
中科院分区：
文献类型：
--
作者：
Hiroki Tsumoto;Yoshihiro Akimoto;Tamao Endo;Yuri Miura.
蛋白质O-GlcNAc化调节多种生物学过程，并且与多种疾病相关。因此，定量蛋白质组学的发展对于了解O-GlcNAc相关疾病的发病机制具有重要意义。我们先前报道了O-GlcNAc酰化肽的选择性富集，其通过新颖的硫醇-炔和硫醇-二硫键交换提供高选择性和有效的释放。在这里，我们描述了一种新的方法，使用初始同量异序标签标记的相对定量，然后富集和β-消除/迈克尔加成与二硫苏糖醇的蛋白质和修饰位点的鉴定。使用模型蛋白质和肽验证该方法。该方法可用于生物样品中O-GlcNAcome的定量分析。
ProteinO-GlcNAcylation regulates various biological processes, and is associated with several diseases. Therefore, the development of quantitative proteomics is important for understanding the mechanisms ofO-GlcNAc-related diseases. We previously reported selective enrichment ofO-GlcNAcylated peptides, which provided high-selectivity and effective release by a novel thiol-alkyne and thiol-disulfide exchange. Here, we describe a new approach using initial isobaric tag labeling for relative quantification followed by enrichment and β-elimination/Michael addition with dithiothreitol for identification of both proteins and modification sites. The approach was validated using model proteins and peptides. This novel strategy could be used for quantitativeO-GlcNAcome of biological samples.