Structure-Guided Design and In-Cell Target Profiling of a Cell-Active Target Engagement Probe for PARP Inhibitors.,ACS Chemical Biology

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Structure-Guided Design and In-Cell Target Profiling of a Cell-Active Target Engagement Probe for PARP Inhibitors.
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2020-02-10 , DOI: 10.1021/acschembio.9b00963
Ryan T Howard ₁ , Paul Hemsley ₂ , Philip Petteruti ₃ , Charlie N Saunders ₁ , Javier A Molina Bermejo ₁ , James S Scott ₂ , Jeffrey W Johannes ₃ , Edward W Tate ₁

Affiliation

Inhibition of the poly(ADP-ribose) polymerase (PARP) family of enzymes has become an attractive therapeutic strategy in oncology and beyond; however, chemical tools to profile PARP engagement in live cells are lacking. Herein, we report the design and application of PARPYnD, the first photoaffinity probe (AfBP) for PARP enzymes based on triple PARP1/2/6 inhibitor AZ9482, which induces multipolar spindle (MPS) formation in breast cancer cells. PARPYnD is a robust tool for profiling PARP1/2 and is used to profile clinical PARP inhibitor olaparib, identifying several novel off-target proteins. Surprisingly, while PARPYnD can enrich recombinant PARP6 spiked into cellular lysates and inhibits PARP6 in cell-free assays, it does not label PARP6 in intact cells. These data highlight an intriguing biomolecular disparity between recombinant and endogenous PARP6. PARPYnD provides a new approach to expand our knowledge of the targets of this class of compounds and the mechanisms of action of PARP inhibitors in cancer.

中文翻译：

PARP抑制剂的细胞活性靶标探针的结构指导设计和细胞内靶标分析。

抑制聚（ADP-核糖）聚合酶（PARP）酶家族已成为肿瘤学及其他领域中有吸引力的治疗策略。但是，缺乏用于分析PARP参与活细胞的化学工具。本文中，我们报道了PARPYnD的设计和应用，PARPYnD是第一个基于三重PARP1 / 2/6抑制剂AZ9482的PARP酶光亲和探针（AfBP），该探针可诱导乳腺癌细胞中多极纺锤体（MPS）的形成。PARPYnD是用于分析PARP1 / 2的强大工具，可用于分析临床PARP抑制剂olaparib，从而鉴定出几种新型的脱靶蛋白。令人惊讶的是，尽管PARPYnD可以富集掺入细胞裂解物中的重组PARP6，并在无细胞测定中抑制PARP6，但它并未在完整细胞中标记PARP6。这些数据突显了重组和内源性PARP6之间令人感兴趣的生物分子差异。PARPYnD提供了一种新方法，可扩展我们对这类化合物的靶标以及PARP抑制剂在癌症中的作用机制的认识。

更新日期：2020-02-11

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