蛋白质组
反褶积
蛋白质组学
计算生物学
吞吐量
工作流程
质谱法
化学
计算机科学
色谱法
生物
生物化学
算法
基因
数据库
电信
无线
作者
Massimiliano Gaetani,Roman A. Zubarev
出处
期刊:Methods in molecular biology
日期:2022-10-01
卷期号:: 91-106
被引量:15
标识
DOI:10.1007/978-1-0716-2624-5_7
摘要
Proteome Integral Solubility Alteration (PISA) is a recently developed mass spectrometry-based, deep proteomics method for unbiased, proteome-wide target deconvolution of ligands, requiring no chemical ligand modification. PISA can be applied to living cells for studying target engagement in vivo or alternatively to protein extracts to identify in vitro ligand-interacting proteins. Here we describe the PISA workflow optimized in our lab. PISA improves the target discovery throughput 10-100 folds compared to the previously used proteomics methods and provides higher statistical significance for target candidates by enabling several biological replicates. Sample multiplexing makes all-in-one analysis of multiple ligands simultaneously possible. PISA dramatically reduces analysis costs, allowing many research questions in need of target deconvolution to be addressed, and unlocks the potential of miniaturizing biological models, including primary cells.
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