RAS-inhibiting biologics identify and probe druggable pockets including an SII-a3 allosteric site
RAS-inhibiting biologics identify and probe druggable pockets including an SII-a3 allosteric site
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RAS 抑制生物制剂识别和探测可药物口袋，包括 SII-a3 变构位点
DOI：
10.1101/2020.06.04.133728
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Haza K
中科院分区：
文献类型：
--
作者：
Haza K
RAS突变是人类癌症中最常见的致癌驱动因素，但临床验证的RAS药理学抑制剂仍然缺乏，因为可药物口袋已被证明难以识别。在这里，我们鉴定了两个ras结合的afferimer蛋白K3和K6，它们抑制核苷酸交换和下游信号通路，具有不同的异构体和突变谱。Affimer K6结合在SI/SII口袋中，而Affimer K3是SII区域的非共价抑制剂，揭示了野生型RAS的一个具有大的，可药物的SII/α3口袋的构象。与RAS结合的定位子和已知的RAS结合小分子之间的竞争性纳米obret证明了利用定位子作为识别药效团的工具的潜力。这项工作强调了使用具有小界面表面的生物制剂来选择看不见的、可药物的构象，并结合药效团识别难以药物的蛋白质的潜力。
RAS mutations are the most common oncogenic drivers across human cancers, but there remains a paucity of clinically-validated pharmacological inhibitors of RAS, as druggable pockets have proven difficult to identify. Here, we identify two RAS-binding Affimer proteins, K3 and K6, that inhibit nucleotide exchange and downstream signaling pathways with distinct isoform and mutant profiles. Affimer K6 binds in the SI/SII pocket, whilst Affimer K3 is a non-covalent inhibitor of the SII region that reveals a conformer of wild-type RAS with a large, druggable SII/α3 pocket. Competitive NanoBRET between the RAS-binding Affimers and known RAS binding small-molecules demonstrates the potential to use Affimers as tools to identify pharmacophores. This work highlights the potential of using biologics with small interface surfaces to select unseen, druggable conformations in conjunction with pharmacophore identification for hard-to-drug proteins.