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在酸性pH下标记TMT克服了有害的过度标记,并提高了的识别率

Rijing Liao¹, Pu You², Kai Weng¹

¹Shanghai Institute of Precision Medicine, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200125, China.

Analytical chemistry

|July 5, 2023

概括

这项研究揭示了Tandem Mass Tags (TMT) 过度标签是由胺催化剂引起的. 一种新的酸性pHTMT标签方法可以防止这种情况,在蛋白质组学中显著增加了和蛋白质的识别.

科学领域:

蛋白质组学是指蛋白质组学
分析化学分析化学
生物化学生物化学

背景情况:

串联质量标签 (TMT) 通过多重化样本来实现高通量蛋白质组学量化.
标准TMT标签的目标是初级氨基,但也可以标记基,降低灵敏度和的识别.
过度标签,特别是氨酸,氨酸和氨酸残留物,危害了基于TMT的蛋白质组分析.

研究的目的:

为了调查TMT过度标签背后的化学机制.
开发一个改进的TMT标签策略,以减轻过度标签.
在定量蛋白质组学中提高和蛋白质识别率.

主要方法:

研究了TMT过度标记的化学性质,通过histidine残留物识别了分子内催化.
开发了一种修改的TMT标签协议,利用酸性pH条件.
使用蛋白质组分析,将新方法与标准TMT标签程序进行比较.

主要成果:

鉴定出以histidine为媒介的分子内催化是TMT过度标记基团的原因.
新的酸性pHTMT标签方法有效地防止了过度标签.
实现了与标准方法相比较的标签效率,同时显著减少了标签过重的.
导致独特的增加33.9%,蛋白质识别的增加20.9%.

结论:

了解TMT覆盖标签的化学机制对于方法开发至关重要.
开发的酸性pHTMT标签方法为定量蛋白质组学提供了显著的改进.
这种优化的TMT方法提高了分析灵敏度和数据质量,使得蛋白质覆盖范围更深.

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