透电荷溶解透与原生蛋白质离子并极大地影响上下序列数据
Daniel A Polasky1, Sugyan M Dixit1, Michael F Keating1
1Department of Chemistry, University of Michigan, 930 North University Avenue, Ann Arbor, Michigan 48109, United States.
Journal of the American Chemical Society
|March 24, 2020
概括
用三甲基 (TMP) 标记蛋白质改变了碎片化途径,从而改善了自上而下的测序. 这种方法通过控制电荷和改善完整蛋白质分析的序列覆盖来增强蛋白质分析.
科学领域:
- 蛋白质组学
- 质谱学
- 生物化学
背景情况:
- 翻译后的修改会产生多样化的蛋白质形式,使与疾病相关的蛋白质组分析复杂化.
- 完整蛋白质的自上而下的测序是有利的,但在实现完整的碎片化方面面临挑战.
研究的目的:
- 开发用于完整蛋白质增强碎片化的新策略,以进行全面的上下测序.
- 研究化学标签对蛋白质离子碎片化途径的影响.
主要方法:
- 蛋白质被标记为三甲基 (TMP) 在氨酸残留物.
- 使用碳胺化学方法封闭酸性侧链.
- 通过分子动力学模拟支持的质谱分析了碎片化途径.
主要成果:
- 通过减少移动质子,TMP标签将碎片化从电荷定向向电荷远程路径.
- 覆盖酸性残留物恢复电荷定向的碎片化,导致更均的序列覆盖.
- 碎片化几乎只针对标记为TMP的氨酸残留物.
- 完整的蛋白质离子具有高电荷溶解能力,涉及螺旋形成等结构变化.
结论:
- 完整蛋白质的化学修饰提供了一种强大的方法来控制从上到下测序的碎片.
- 了解气相蛋白离子中的电荷溶解对于推进全面的蛋白质分析至关重要.
- 这些发现为影响上下测序效率的物理力量提供了洞察力.
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