Griffith Institute for Drug Discovery, Griffith University, 46 Don Young Road, Nathan, QLD 4111, Australia.
Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
ChemMedChem. 2021 Jul 20;16(14):2206-2210. doi: 10.1002/cmdc.202100113. Epub 2021 May 4.
PROteolysis TArgeting Chimeras (PROTACs) promote the degradation, rather than inhibition, of a drug target as a mechanism for therapeutic treatment. Bifunctional PROTAC molecules allow simultaneous binding of both the target protein and an E3-Ubiquitin ligase, bringing the two proteins into close spatial proximity to allow ubiquitinylation and degradation of the target protein via the cell's endogenous protein degradation pathway. We utilized native mass spectrometry (MS) to study the ternary complexes promoted by the previously reported PROTAC GNE-987 between Brd4 bromodomains 1 and 2, and Von Hippel Lindeau E3-Ubiquitin Ligase. Native MS at high resolution allowed us to measure ternary complex formation as a function of PROTAC concentration to provide a measure of complex affinity and stability, whilst simultaneously measuring other intermediate protein species. Native MS provides a high-throughput, low sample consumption, direct screening method to measure ternary complexes for PROTAC development.
蛋白水解靶向嵌合体（PROTACs）通过促进药物靶标的降解而不是抑制来发挥作用，这是一种治疗方法的机制。双功能 PROTAC 分子允许同时结合靶蛋白和 E3-泛素连接酶，使两种蛋白质紧密接近，从而通过细胞内的内源性蛋白质降解途径使靶蛋白泛素化和降解。我们利用天然质谱（MS）研究了先前报道的 PROTAC GNE-987 促进的 Brd4 溴结构域 1 和 2 与 Von Hippel Lindeau E3-泛素连接酶之间的三元复合物。高分辨率的天然 MS 使我们能够测量三元复合物的形成作为 PROTAC 浓度的函数，以提供复合物亲和力和稳定性的度量，同时测量其他中间蛋白质物种。天然 MS 提供了一种高通量、低样品消耗、直接筛选方法，用于测量 PROTAC 开发中的三元复合物。
