蛋白质组学
化学
蛋白质沉淀
质谱法
差速离心
分馏
色谱法
降水
肽
自下而上蛋白质组学
蛋白质组
溶解度
计算生物学
串联质谱法
生物化学
蛋白质质谱法
生物
物理
有机化学
气象学
基因
作者
Antonio Frederico Michel Pinto,Jolene K. Diedrich,James J. Moresco,John R. Yates
标识
DOI:10.1021/jasms.3c00182
摘要
Differential precipitation of proteins (DiffPOP) is a simple technique for fractionating complex protein mixtures. Using stepwise addition of acidified methanol, ten distinct subsets of proteins can be selectively precipitated by centrifugation and identified by mass spectrometry-based proteomics. We have previously shown that the ability of a protein to resist precipitation can be altered by drug binding, which enabled us to identify a novel drug–target interaction. Here, we show that the addition of DiffPOP to a standard LC–MS proteomics workflow results in a three-dimensional separation of peptides that increases protein coverage and peptide identifications. Importantly, DiffPOP reveals solubility differences between proteoforms, potentially providing valuable insights that are typically lost in bottom-up proteomics.
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