使用可点击的氨氧基探针检测蛋白质ADP-核糖基化
Detecting Protein ADP-Ribosylation Using a Clickable Aminooxy Probe.
作者信息
Morgan Rory K, Cohen Michael S
机构信息
Program in Chemical Biology, Department of Physiology and Pharmacology, Oregon Health and Science University, 3181 SW Sam Jackson Park Rd, #L334, Biomedical Research Building, Rm 621, Portland, OR, 97239-3098, USA.
出版信息
Methods Mol Biol. 2017;1608:71-77. doi: 10.1007/978-1-4939-6993-7_6.
Abstract
ADP-ribosylation, a posttranslational modification catalyzed by a family of enzymes known as poly(ADP-ribose) polymerases (PARPs, 17 in humans), regulates diverse cellular processes. To aid in understanding the functions of ADP-ribosylation in cells, we developed a clickable aminooxy probe, AO-alkyne, which detects ADP-ribosylation of acidic amino acids. AO-alkyne can be used to detect auto-ADP-ribosylation of PARP10 in cells following Cu-catalyzed click conjugation to an azide reporter. This method can be extended to other PARP family members that catalyze ADP-ribosylation on acidic amino acids, providing a convenient and direct readout of PARP activity in cells.
摘要
ADP核糖基化是一种由称为聚（ADP-核糖）聚合酶（PARP，人类中有17种）的酶家族催化的翻译后修饰，可调节多种细胞过程。为了帮助理解ADP核糖基化在细胞中的功能，我们开发了一种可点击的氨氧基探针AO-炔烃，它可以检测酸性氨基酸的ADP核糖基化。在铜催化下与叠氮化物报告基团进行点击共轭后，AO-炔烃可用于检测细胞中PARP10的自ADP核糖基化。该方法可以扩展到其他在酸性氨基酸上催化ADP核糖基化的PARP家族成员，为细胞中PARP活性提供一种方便且直接的检测方法。
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