通过使用MALDI-MS和MS/MS,通过maleimide化学制备的异性交联的分化
Luis Javier González1, Satomy Pousa1, Hironobu Hojo2
1Mass Spectrometry Laboratory, Department of Proteomics, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Rapid communications in mass spectrometry : RCM
|December 21, 2023
概括
生物结合物中的提奥苏胺结合剂可以转化为水解或 thiazine 形式. 像MALDI-MS和MALDI-MS/MS这样的质谱技术可以有效地区分这些形式的囊烯酸.
科学领域:
- 生物结合化学 生物结合化学
- 分析化学 分析化学
- 质谱测量质量谱测量
背景情况:
- 硫胺连接器是生物结合物合成中常见的工具.
- 这种连接器易于水解,形成水解和 thiazine 变种.
- 氨酸形式在与氨基酸的结合物中很常见.
研究的目的:
- 为了研究在生物结合物中 thiosuccinimide 结合物的稳定性.
- 使用质谱学来区分硫胺,水解和硫胺的形式.
- 为了描述这些等体物种的碎片化模式.
主要方法:
- 合成四个交叉链接的类与thio-succinimide链接剂.
- 在中性或基本pH下进行化,以产生水解和 thiazine 类型.
- 使用逆相高性能液态染色学 (RP-HPLC) 进行净化.
- 使用矩阵辅助激光脱/电离质谱法 (MALDI-MS),MALDI-MS/MS和紫外线光解离法进行差异化.
主要成果:
- 在中性/基本条件下,氨基联体中的氨基胺结合剂在很大程度上转化为水解或氨酸形式.
- 马尔迪-MS区分了两种形式:硫胺和水解产品在减少性裂变时产生构成,而氨酸形式没有产生碎片离子.
- 苏胺胺形式的MALDI-MS/MS显示了1,4-消除碎片,与减少裂解碎片不同.
- 氨酸形式显示出新形成的胺键裂变的碎片化.
结论:
- 马尔迪-MS和马尔迪-MS/MS可以区分酸胺,化和 thiazine 形式的交联.
- 特定的碎片化模式,包括1,4-消除的thiosuccinimide和转循环裂变的thiazine,援助差异化.
- 这些质谱法对于分析含有潜在不稳定的硫胺结合剂的生物结合物至关重要.
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