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首页> 外文期刊>The FEBS journal >Identification of candidate substrates for poly(ADP-ribose) polymerase-2 (PARP2) in the absence of DNA damage using high-density protein microarrays
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Identification of candidate substrates for poly(ADP-ribose) polymerase-2 (PARP2) in the absence of DNA damage using high-density protein microarrays

机译：使用高密度蛋白微阵列在没有DNA损伤的情况下鉴定聚（ADP-核糖）聚合酶2（PARP2）的候选底物

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摘要

Poly(ADP-ribose) polymerase-2 (PARP2) belongs to the ADP-ribosyltransferase family of enzymes that catalyze the addition of ADP-ribose units to acceptor proteins, thus affecting many diverse cellular processes. In particular, PARP2 shares with PARP1 and, as recently highlighted, PARP3 the sole property of being catalytically activated by DNA-strand breaks, implying key downstream functions in the cellular response to DNA damage for both enzymes. However, evidence from several studies suggests unique functions for PARP2 in additional processes, possibly mediated through its basal, DNA-damage unstimulated ADP-ribosylating activity. Here, we describe the development and application of a protein microarray-based approach tailored to identify proteins that are ADP-ribosylated by PARP2 in the absence of DNA damage mimetics and might thus represent useful entry points to the exploration of novel PARP2 functions. Several candidate substrates for PARP2 were identified and global hit enrichment analysis showed a clear enrichment in translation initiation and RNA helicase molecular functions. In addition, the top scoring candidates FK506-binding protein 3 and SH3 and cysteine-rich domain-containing protein 1 were selected and confirmed in a complementary assay format as substrates for unstimulated PARP2.
机译：聚（ADP-核糖）聚合酶-2（PARP2）属于酶的ADP-核糖基转移酶家族，可催化​​将ADP-核糖单元添加到受体蛋白上，从而影响许多不同的细胞过程。特别是，PARP2与PARP1共享，并且最近强调指出，PARP3的唯一特性是被DNA链断裂催化激活，这暗示了细胞对两种酶对DNA损伤的反应中的关键下游功能。但是，一些研究的证据表明，PARP2在其他过程中具有独特的功能，可能是通过其基础的，DNA损伤的未刺激的ADP-核糖基化活性介导的。在这里，我们描述了一种基于蛋白质微阵列的方法的开发和应用，该方法旨在识别在没有DNA损伤模拟物的情况下被PARP2 ADP核糖基化的蛋白质，因此可能代表了探索新型PARP2功能的有用切入点。鉴定了PARP2的几种候选底物，全局命中富集分析表明翻译起始和RNA解旋酶分子功能明显富集。另外，选择得分最高的候选FK506结合蛋白3和SH3以及富含半胱氨酸的域的蛋白1，并以互补测定形式确认为未刺激的PARP2的底物。

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