将自上而下的质谱扩展到质量大于200千多的蛋白质
Xuemei Han1, Mi Jin, Kathrin Breuker
1Department of Chemistry and Chemical Biology, Baker Laboratory, Cornell University, Ithaca, NY 14853, USA.
概括
顶向下质谱现在通过克服以前的尺寸限制来分析更大的蛋白质. 这种先进的方法提供了高度特定的分子和碎片离子数据,用于详细的蛋白质表征.
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:
- 传统的蛋白质分析依赖于质量数据,这比分子和碎片离子数据更不具体.
- 顶向下的质谱学提供了更高的特异性,但仅限于较小的蛋白质 (约. 50 kDa) 的时间.
研究的目的:
- 为了将上下质谱的适用性扩展到更大的蛋白质.
- 为了增强蛋白质序列和翻译后修改的特征.
主要方法:
- 使用电喷剂添加剂,加热蒸发和分离非共价和共价键解离技术.
- 应用了这些方法来克服上下质谱学的大约500余分离限制.
主要成果:
- 在一个1314-残留 (144-kD) 蛋白质中成功切割了287个间残留键.
- 在1714残留 (200-kD) 蛋白中,在27种囊蛋白中的8种中确定了以前未知的二硫化键.
- 修正了两个大型蛋白质的序列预测,包括一个具有2153个残留物 (229 kD) 的蛋白质.
结论:
- 增强的自上而下的质谱方法显著扩大了可以分析的蛋白质大小范围.
- 这种方法为蛋白质表征提供了更具体和更全面的方法,包括序列和二硫化键分析.
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