Proteomics approaches to identify mono-(ADP-ribosyl)ated and poly(ADP-ribosyl)ated proteins.
Proteomics approaches to identify mono-(ADP-ribosyl)ated and poly(ADP-ribosyl)ated proteins.
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DOI：
10.1002/pmic.201400217
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发表时间：
2015-01
期刊：
影响因子：
3.4
通讯作者：
Leung AK
中科院分区：
文献类型：
--
作者：
Vivelo CA;Leung AK
ADP-核糖基化是指在蛋白质底物上添加一个或多个ADP-核糖单元，这种蛋白质修饰涉及各种细胞过程，包括DNA损伤修复、RNA代谢、转录和细胞周期调控。本文综述了大规模蛋白质组学研究的汇编，确定ADP-核糖基化蛋白质及其相关蛋白质的质谱使用各种富集策略。一些方法，如使用聚（ADP-核糖）特异性抗体和硼酸盐亲和色谱法和NAD+类似物，已经使用了几十年，而其他方法，如使用蛋白质微阵列和结合ADP-核糖部分（如大结构域）的重组蛋白，只是最近才开发出来。每种方法的优点和缺点，以及这些方法是否是具体的识别单（ADP-核糖基）和聚（ADP-核糖基）的蛋白质将进行讨论。最后，由于聚（ADP-核糖）在长度上是异质的，因此难以获得与修饰位点相关的质量特征。本文综述了几种降低聚合物链长异质性的方法。
ADP-ribosylation refers to the addition of one or more ADP-ribose units onto protein substrates and this protein modification has been implicated in various cellular processes including DNA damage repair, RNA metabolism, transcription and cell cycle regulation. This review focuses on a compilation of large-scale proteomics studies that identify ADP-ribosylated proteins and their associated proteins by mass spectrometry using a variety of enrichment strategies. Some methods, such as the use of a poly(ADP-ribose)-specific antibody and boronate affinity chromatography and NAD+ analogues, have been employed for decades while others, such as the use of protein microarrays and recombinant proteins that bind ADP-ribose moieties (such as macrodomains), have only recently been developed. The advantages and disadvantages of each method and whether these methods are specific for identifying mono(ADP-ribosyl)ated and poly(ADP-ribosyl)ated proteins will be discussed. Lastly, since poly(ADP-ribose) is heterogeneous in length, it has been difficult to attain a mass signature associated with the modification sites. Several strategies on how to reduce polymer chain length heterogeneity for site identification will be reviewed.