化学
溶解
肽
多路复用
计算生物学
仿形(计算机编程)
小分子
蛋白质组学
生物化学
计算机科学
电信
生物
基因
操作系统
作者
He Zhu,Scott B. Ficarro,William M. Alexander,Laura E Fleming,Guillaume Adelmant,Tinghu Zhang,Matthew Willetts,Jens Decker,Sven Brehmer,Michael Krause,Michael P. East,Nathanael S. Gray,Gary L. Johnson,Gary Kruppa,Jarrod A. Marto
出处
期刊:Analytical Chemistry
[American Chemical Society]
日期:2021-10-04
卷期号:93 (41): 13791-13799
被引量:17
标识
DOI:10.1021/acs.analchem.1c02349
摘要
Parallel reaction monitoring (PRM) has emerged as a popular approach for targeted protein quantification. With high ion utilization efficiency and first-in-class acquisition speed, the timsTOF Pro provides a powerful platform for PRM analysis. However, sporadic chromatographic drift in peptide retention time represents a fundamental limitation for the reproducible multiplexing of targets across PRM acquisitions. Here, we present PRM-LIVE, an extensible, Python-based acquisition engine for the timsTOF Pro, which dynamically adjusts detection windows for reproducible target scheduling. In this initial implementation, we used iRT peptides as retention time standards and demonstrated reproducible detection and quantification of 1857 tryptic peptides from the cell lysate in a 60 min PRM-LIVE acquisition. As an application in functional proteomics, we use PRM-LIVE in an activity-based protein profiling platform to assess binding selectivity of small-molecule inhibitors against 220 endogenous human kinases.
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