首页> 外文期刊>Nature >USP7 small-molecule inhibitors interfere with ubiquitin binding
【24h】

USP7 small-molecule inhibitors interfere with ubiquitin binding

机译:USP7小分子抑制剂干扰泛素结合

获取原文
获取原文并翻译 | 示例
       

摘要

The ubiquitin system regulates essential cellular processes in eukaryotes. Ubiquitin is ligated to substrate proteins as monomers or chains and the topology of ubiquitin modifications regulates substrate interactions with specific proteins. Thus ubiquitination directs a variety of substrate fates including proteasomal degradation(1). Deubiquitinase enzymes cleave ubiquitin from substrates and are implicated in disease(2); for example, ubiquitin-specific protease-7 (USP7) regulates stability of the p53 tumour suppressor and other proteins critical for tumour cell survival(3). However, developing selective deubiquitinase inhibitors has been challenging(4) and no co-crystal structures have been solved with small-molecule inhibitors. Here, using nuclear magnetic resonance-based screening and structure-based design, we describe the development of selective USP7 inhibitors GNE-6640 and GNE-6776. These compounds induce tumour cell death and enhance cytotoxicity with chemotherapeutic agents and targeted compounds, including PIM kinase inhibitors. Structural studies reveal that GNE-6640 and GNE-6776 non-covalently target USP7 12 angstrom distant from the catalytic cysteine. The compounds attenuate ubiquitin binding and thus inhibit USP7 deubiquitinase activity. GNE-6640 and GNE-6776 interact with acidic residues that mediate hydrogen-bond interactions with the ubiquitin Lys48 side chain5, suggesting that USP7 preferentially interacts with and cleaves ubiquitin moieties that have free Lys48 side chains. We investigated this idea by engineering di-ubiquitin chains containing differential proximal and distal isotopic labels and measuring USP7 binding by nuclear magnetic resonance. This preferential binding protracted the depolymerization kinetics of Lys48-linked ubiquitin chains relative to Lys63-linked chains. In summary, engineering compounds that inhibit USP7 activity by attenuating ubiquitin binding suggests opportunities for developing other deubiquitinase inhibitors and may be a strategy more broadly applicable to inhibiting proteins that require ubiquitin binding for full functional activity.
机译:泛素系统调节真核生物中必需的细胞过程。泛素作为单体或链与底物蛋白质连接,泛素修饰的拓扑结构调节底物与特定蛋白质的相互作用。因此泛素化指导了多种底物的命运,包括蛋白酶体降解(1)。去泛素酶从底物中切割泛素,并与疾病有关(2);例如,泛素特异性蛋白酶7(USP7)调节p53肿瘤抑制因子和其他对肿瘤细胞存活至关重要的蛋白质的稳定性(3)。然而,开发选择性的去泛素酶抑制剂具有挑战性(4),并且小分子抑制剂还没有解决共晶结构。在这里,使用基于核磁共振的筛选和基于结构的设计,我们描述了选择性USP7抑制剂GNE-6640和GNE-6776的开发。这些化合物可通过化学治疗剂和靶向化合物(包括PIM激酶抑制剂)诱导肿瘤细胞死亡并增强细胞毒性。结构研究表明,GNE-6640和GNE-6776非共价地靶向距催化半胱氨酸12埃的USP7。这些化合物减弱了泛素结合并因此抑制了USP7去泛素酶活性。 GNE-6640和GNE-6776与酸性残基相互作用,该残基介导与泛素Lys48侧链的氢键相互作用5,这表明USP7优先与具有游离Lys48侧链的泛素部分相互作用并切割。我们通过工程化包含差异性近端和远端同位素标记的双泛素链并通过核磁共振测量USP7结合来研究此想法。相对于Lys63连接的链,这种优先结合延长了Lys48连接的泛素链的解聚动力学。总而言之,通过减弱泛素结合来抑制USP7活性的工程化合物提示开发其他去泛素酶抑制剂的机会,并且可能是更广泛适用于抑制需要泛素结合才能发挥全部功能活性的蛋白质的策略。

著录项

  • 来源
    《Nature》 |2017年第7677期|534-538|共5页
  • 作者单位

    Genentech Inc, Dept Discovery Oncol, San Francisco, CA 94080 USA|Genentech Inc, Dept Early Discovery Biochem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Struct Biol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Struct Biol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Chem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Biochem & Cellular Pharmacol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Biochem & Cellular Pharmacol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Biochem & Cellular Pharmacol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Oncol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Oncol, San Francisco, CA 94080 USA|Genentech Inc, Dept Early Discovery Biochem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Oncol, San Francisco, CA 94080 USA|Genentech Inc, Dept Early Discovery Biochem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Oncol, San Francisco, CA 94080 USA|Genentech Inc, Dept Early Discovery Biochem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Oncol, San Francisco, CA 94080 USA|Genentech Inc, Dept Early Discovery Biochem, San Francisco, CA 94080 USA;

    Univ Dundee, Sch Life Sci, MRC Prot Phosphorylat & Ubiquitylat Unit, Dundee DD1 5EH, Scotland;

    Univ Dundee, Sch Life Sci, MRC Prot Phosphorylat & Ubiquitylat Unit, Dundee DD1 5EH, Scotland;

    Newcastle Univ, Inst Cell & Mol Biosci, Newcastle Upon Tyne NE2 1HH, Tyne & Wear, England;

    Genentech Inc, Dept Prot Chem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Prot Chem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Microchem Prote & Lipid, San Francisco, CA 94080 USA;

    Boston Biochem, 840 Mem Dr, Cambridge, MA 02139 USA;

    Boston Biochem, 840 Mem Dr, Cambridge, MA 02139 USA;

    Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA|Univ Calif San Francisco, Small Mol Discovery Ctr, San Francisco, CA 94143 USA;

    Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA|Univ Calif San Francisco, Small Mol Discovery Ctr, San Francisco, CA 94143 USA;

    Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA|Univ Calif San Francisco, Small Mol Discovery Ctr, San Francisco, CA 94143 USA;

    Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA|Univ Calif San Francisco, Small Mol Discovery Ctr, San Francisco, CA 94143 USA;

    Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA|Univ Calif San Francisco, Small Mol Discovery Ctr, San Francisco, CA 94143 USA;

    Genentech Inc, Dept Discovery Chem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Microchem Prote & Lipid, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Res Pathol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Bioinformat & Computat Biol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Bioinformat & Computat Biol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Bioinformat & Computat Biol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Translat Oncol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Oncol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Bioinformat & Computat Biol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Prot Chem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Chem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Chem, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Biochem & Cellular Pharmacol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Chem, San Francisco, CA 94080 USA;

    Boston Biochem, 840 Mem Dr, Cambridge, MA 02139 USA;

    Genentech Inc, Dept Biochem & Cellular Pharmacol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Struct Biol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Struct Biol, San Francisco, CA 94080 USA;

    Genentech Inc, Dept Discovery Oncol, San Francisco, CA 94080 USA|Genentech Inc, Dept Early Discovery Biochem, San Francisco, CA 94080 USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

  • 入库时间 2022-08-18 02:51:56

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号