紫外线光解离允许对紫外线进行全面的描述
Amanda Helms1, Edwin E Escobar1,2, Saulius Vainauskas2
1Department of Chemistry, The University of Texas at Austin, Austin, Texas 78712, United States.
Analytical chemistry
|June 8, 2023
概括
描述O-糖,包括它们的位点和糖,是具有挑战性的. 使用O-糖蛋白酶IMPa和紫外光解离 (UVPD) 的新策略成功地在复杂的糖蛋白上映射了多个O-糖体和糖体.
科学领域:
- 生物化学 生物化学
- 蛋白质组学是指蛋白质组学.
- 葡萄糖科学 (Glycoscience) 是一种科学.
背景情况:
- 对O-糖的表征至关重要,但在技术上具有挑战性,特别是对于具有异质糖结构的多糖化.
- 现有的方法难以对O-甘氨酸进行全面的分析,包括标识,甘氨酸位定位和甘氨酸映射.
- 紫外线光解离 (UVPD) 显示出局部化多个翻译后修饰和特征糖的潜力.
研究的目的:
- 开发和验证一个完整的O-糖特性策略.
- 评估将O-葡萄糖蛋白酶IMPa与HCD触发的UVPD结合用于分析复杂的葡萄糖蛋白的疗效.
- 为了比较UVPD与HCD和EThcD的性能,用于O-糖位定位和糖分析.
主要方法:
- 使用的O-糖蛋白酶IMPa用于糖蛋白的酶性消化.
- 使用HCD触发的UVPD进行糖的碎片化和分析.
- 分析了三种葡萄糖蛋白,包括乙素,以评估组合方法.
主要成果:
- 在单个糖上成功局部化了多个相邻或近邻的O-糖.
- 在etanercept上确定了一种新的O-糖在血清218 (S218) 中.
- 从etanercept的多糖基化中特征了九种不同的甘油形式.
- 与HCD和EThcD相比,UVPD的性能用于O-糖位定位和糖/糖的表征.
结论:
- 结合的O-糖蛋白酶IMPA和HCD触发的UVPD策略使O-糖的全面表征成为可能.
- 这种方法克服了分析多糖基化和异质甘氨酸结构的挑战.
- 在葡萄糖保护学中,UVPD为O-葡萄糖酸盐局部化和葡萄糖酸盐分析提供了强大的替代方案.
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