在快速获取飞行时间质谱仪上使用电子激活解离碎片定位和量化蛋白质的翻译后修饰
Joanna Bons1, Christie L Hunter2, Rita Chupalov3
1Buck Institute for Research on Aging, Novato, California 94947, United States.
Journal of the American Society for Mass Spectrometry
|September 11, 2023
概括
电子激活解离 (EAD) 与碰撞诱导解离 (CID) 相比,为表征可变的翻译后修饰 (PTM) 提供了优越的碎片化. 这种新方法提高了PTM位点定位和量化准确性,提高了蛋白质组分析的灵敏度.
科学领域:
- 蛋白质组学是指蛋白质组学.
- 质谱测量质量谱测量
- 生物化学 生物化学
背景情况:
- 翻译后修饰 (PTM) 对细胞过程至关重要,但通过蛋白质组学进行分析具有挑战性.
- 像碰撞诱导解离 (CID) 这样的常见碎片化方法可以导致中性损失和不完整的PTM序列.
研究的目的:
- 评估电子激活解离 (EAD) 作为PTM分析的替代碎片化技术.
- 为了比较EAD与CID的性能,以表征,定位和量化具有可变修饰的.
主要方法:
- 使用有针对性的并行反应监测 (PRM) 分析化,化,化和乙化合成.
- 在快速扫描四极飞行时间 (QqTOF) 质谱仪上对EAD和CID碎片化的比较.
- 利用一种新的陷和释放技术来增强EAD的灵敏度和工作周期.
主要成果:
- EAD有效地保留了包括马洛尼尔组在内的可变性修饰物,防止中性损失.
- EAD提供了增强的序覆盖范围和更强的PTM位点定位碎片离子.
- EAD工作流显示出高可重现性 (CVs约为2-7%),线性性和量化准确性,MS/MS灵敏度增加了6-11倍.
结论:
- EAD提供了一种强大的替代碎片化方法,用于自信的PTM表征和量化.
- EAD和陷释放技术的组合显著提高了PTM分析中的灵敏度和分析性能.
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