LC-MS/MS 蛋白质形状分析暴露了线粒体中的细胞染色体c 过氧化酶自氧化和其血液中的功能性重要洞跳跃
Meena Kathiresan1, Ann M English1
1Quebec Network for Research on Protein Function, Structure and Engineering (PROTEO), and Department of Chemistry and Biochemistry , Concordia University , Montreal , QC H4B 1R6 , Canada.
Journal of the American Chemical Society
|August 28, 2018
概括
细胞染色体c过氧化酶 (Ccp1) 蛋白质形式通过基因转移发生氧化,从而显示出新的功能和基质结合位. 这一过程,特别是血标记,扩大了Ccp1
科学领域:
- 生物化学和分子生物学
- 氧化还原酶的功能
- 蛋白质组学
背景情况:
- 细胞染色体c过氧化酶 (Ccp1) 是一个关键的线粒体酶,参与呼吸.
- 氧化应激可以导致蛋白质的修饰,改变它们的功能.
- 了解蛋白质的修饰是解读细胞过程的关键.
研究的目的:
- 研究酵母线粒体中Ccp1蛋白质的氧化变化.
- 阐明Ccp1氧化的机制和功能后果.
- 确定Ccp1的新基质结合点和细胞作用.
主要方法:
- 液体染色学-双重质谱 (LC-MS/MS) 用于蛋白质形状分析.
- 使用重组Ccp1和过氧化 (H2O2) 的体外氧化试验.
- 对外线粒体Ccp1蛋白形态的表征.
主要成果:
- 在Ccp1蛋白形中,LC-MS/MS显示了Met,Trp和Tyr残留的大量氧化.
- 氧化变化与血红素转移 (洞跳转) 一致.
- 穿孔跳跃促进血红素化,氧化H175,并识别新的基质结合位点,包括谷氨.
结论:
- 线粒体中Ccp1的脱离氧化和减少导致新型蛋白质.
- 穿孔跳跃扩展了Ccp1的生理功能,超出了电子转移,包括转移用于酶成熟.
- 通过LC-MS/MS进行蛋白形状分析是研究氧化还原酶细胞作用的强大工具.
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