pSILAC mass spectrometry reveals ZFP91 as IMiD-dependent substrate of the CRL4(CRBN) ubiquitin ligase.
pSILAC mass spectrometry reveals ZFP91 as IMiD-dependent substrate of the CRL4(CRBN) ubiquitin ligase.
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DOI：
10.1038/ncomms15398
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发表时间：
2017-05-22
影响因子：
16.6
通讯作者：
Fischer ES
中科院分区：
文献类型：
--
作者：
An J;Ponthier CM;Sack R;Seebacher J;Stadler MB;Donovan KA;Fischer ES
沙利度胺及其衍生物来那度胺和泊马度胺（IMiDs）是治疗恶性血液病的有效药物。研究表明，IMiDs赋予CUL4-RBX1-DDB1-CRBN （CRL4CRBN）泛素连接酶功能获得特性，使其能够结合、泛素化和降解IKZF1、IKZF3和CSNK1A1等关键治疗靶点。虽然这些底物已被认为是多发性骨髓瘤（MM）和5q缺失相关骨髓增生异常综合征（del(5q)-MDS）的疗效靶点，但可能存在其他靶点。使用基于脉冲追踪SILAC质谱的蛋白质组学方法，我们证明来那度胺诱导ZFP91的泛素化和降解。我们确定ZFP91是一个真正的依赖于imid的CRL4CRBN底物，并进一步证明ZFP91含有一个锌指（ZnF）基序，与IKZF1/3 ZnF相关，对依赖于imid的CRBN结合至关重要。这些研究结果表明，基于单时间点脉冲追踪SILAC质谱的蛋白质组学（pSILAC MS）是一种敏感的靶向鉴定诱导选择性蛋白质降解的小分子的方法。靶向治疗相关蛋白降解是药物发现的一个新兴范例。在这里，作者描述了一种基于敏感脉冲SILAC质谱的蛋白质组学方法，该方法在单时间点实验中报告了药物治疗后蛋白质稳定性的全局变化。
Thalidomide and its derivatives lenalidomide and pomalidomide (IMiDs) are effective treatments of haematologic malignancies. It was shown that IMiDs impart gain-of-function properties to the CUL4-RBX1-DDB1-CRBN (CRL4CRBN) ubiquitin ligase that enable binding, ubiquitination and degradation of key therapeutic targets such as IKZF1, IKZF3 and CSNK1A1. While these substrates have been implicated as efficacy targets in multiple myeloma (MM) and 5q deletion associated myelodysplastic syndrome (del(5q)-MDS), other targets likely exist. Using a pulse-chase SILAC mass spectrometry-based proteomics approach, we demonstrate that lenalidomide induces the ubiquitination and degradation of ZFP91. We establish ZFP91 as a bona fide IMiD-dependent CRL4CRBN substrate and further show that ZFP91 harbours a zinc finger (ZnF) motif, related to the IKZF1/3 ZnF, critical for IMiD-dependent CRBN binding. These findings demonstrate that single time point pulse-chase SILAC mass spectrometry-based proteomics (pSILAC MS) is a sensitive approach for target identification of small molecules inducing selective protein degradation. Targeting therapeutically-relevant proteins for degradation is an emerging paradigm in drug discovery. Here the authors describe a sensitive pulse SILAC mass spectrometry-based proteomics approach that reports global changes in protein stability following drug treatment in a single time point experiment.