Enrichment of O-GlcNAc-modified peptides using novel thiol-alkyne and thiol-disulfide exchange.
Enrichment of O-GlcNAc-modified peptides using novel thiol-alkyne and thiol-disulfide exchange.
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DOI：
10.1016/j.bmcl.2015.04.082
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发表时间：
2015-07
影响因子：
2.7
通讯作者：
H. Tsumoto;Daisuke Ogasawara;N. Hashii;Takayoshi Suzuki;Y. Akimoto;T. Endo;Y. Miura
中科院分区：
文献类型：
--
作者：
H. Tsumoto;Daisuke Ogasawara;N. Hashii;Takayoshi Suzuki;Y. Akimoto;T. Endo;Y. Miura
我们开发了一种选择性富集o -连接β- n -乙酰氨基葡萄糖（O-GlcNAc）修饰肽的方法，该方法使用新合成的硫醇-炔和硫醇-二硫交换。首先，o - glnaac修饰的肽被酶标记为含有叠氮化物的GalNAc类似物。然后，通过铜(I)催化叠氮化物-炔烃环加成反应叠氮化物部分与巯基炔烃反应。巯基修饰肽通过巯基-二硫交换被巯基反应树脂富集。从α-晶体蛋白色氨酸中选择性分离出至少500 fmol的o - glcnac修饰肽，并进行质谱检测。这种新的富集策略可用于生物样品的o - glcnacome分析。
We have developed a selective method for the enrichment ofO-linked β-N-acetylglucosamine (O-GlcNAc)-modified peptides, which uses a newly synthesized thiol-alkyne and a thiol-disulfide exchange. First,O-GlcNAc-modified peptides were enzymatically labeled with an azide-containing GalNAc analog. Then, the azide moiety was reacted with thiol-alkyne through a copper(I)-catalyzed azide-alkyne cycloaddition. The thiol-modified peptides were enriched with thiol-reactive resin through a thiol-disulfide exchange. At least 500 fmol ofO-GlcNAc-modified peptides was selectively isolated from α-crystallin tryptic peptides and detected by mass spectrometry. This novel enrichment strategy could be used forO-GlcNAcome analysis of biological samples.