在自下而上的蛋白质实验中对翻译后和化学修饰的染色学行为汇编
Quinn Neale1, Alexandre Prefontaine1, Taylor Battellino1
1Department of Chemistry, University of Manitoba, 360 Parker Building, Winnipeg R3T 2N2, Manitoba, Canada.
Analytical chemistry
|September 22, 2023
概括
这项研究量化了70多种翻译后修饰的染色学行为. 了解这些疏水性转移有助于预测蛋白质组学中的性保留.
科学领域:
- 蛋白质组学是指蛋白质组学.
- 分析化学 分析化学
- 生物化学 生化学
背景情况:
- 翻译后修改 (PTMs) 显著改变的特性.
- 准确预测保留对于蛋白质组数据分析至关重要.
- 现有的方法缺乏关于各种PTM的染色学影响的全面数据.
研究的目的:
- 系统地评估超过70种翻译后和化学修饰的的染色学行为.
- 为了建立基修饰及其保留时间之间的定量关系.
- 为改善蛋白质组学中类保留的预测模型提供基础.
主要方法:
- 在标准自下而上的蛋白质条件下 (C18,0.1%的酸) 测量修饰/非修饰对的保留特性.
- 通过自发降解,化学/酶处理,合成和非正规氨基酸结合生成对.
- 使用外部数据对合成和酶诱导素的验证.
- 使用疏水性指数 (HI,% ACN) 和平均保留转移 (ΔHI) 的分析.
主要成果:
- 每个修饰都表现出独特的色谱行为,其特点是特定的斜率和截面值,当绘制修饰与非修饰HI时.
- 这些相关性取决于修饰的化学特性 (疏水性,大小,pKa,位置).
- 疏水性指数 (HI) 和保留转移 (ΔHI) 提供了对修改效应的定量衡量.
结论:
- 该研究为广泛的基修饰提供了对色谱行为的系统评估.
- 构成依赖的相关性为将PTM效应纳入保留预测模型提供了基础.
- 开发依赖序列的算法是必要的,以便更准确地预测修改诱导的保留转移.
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