We developed a method for comprehensively profiling drug‐protein interactions using micro‐column affinity purification (AP) combined with label‐free quantitative (LFQ) proteomics as well as the statistical and bioinformatics analysis. FK506 was used as the experimental model for proof of concept. The true interacting proteins were distinguished from the background proteins by their fold changes of FLQ intensities combined with p‐values. Totally 116 FK506 interacting proteins including 5 known target proteins were identified. The method was validated by using the LFQ intensity of the endogenous known drug targets together with statistical analysis. We demonstrated that the micro‐column‐based affinity purification in combination with LFQ proteomics provides a highly reproducible and robust approach for profiling drug‐protein interactions.

中文翻译：

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通过微柱亲和纯化结合无标记定量蛋白质组学分析药物-蛋白质相互作用†

我们开发了一种使用微柱亲和纯化（AP）结合无标记定量（LFQ）蛋白质组学以及统计和生物信息学分析全面分析药物-蛋白质相互作用的方法。FK506用作概念验证的实验模型。真正的相互作用蛋白与背景蛋白的区别在于其FLQ强度与p的倍数变化值。共鉴定出116种FK506相互作用蛋白，包括5种已知靶蛋白。该方法通过使用内源性已知药物靶标的LFQ强度和统计分析进行了验证。我们证明了基于微柱的亲和纯化与LFQ蛋白质组学相结合，为分析药物-蛋白质相互作用提供了一种高度可重复且强大的方法。