VanLinden Magali R, Niere Marc, Nikiforov Andrey A, Ziegler Mathias, Dölle Christian
Department of Molecular Biology, University of Bergen, Thormøhlensgate 55, 5008, Bergen, Norway.
Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia.
Methods Mol Biol. 2017;1608:45-56. doi: 10.1007/978-1-4939-6993-7_4.
Nicotinamide adenine dinucleotide (NAD) is vital to many cellular processes and is distributed between distinct subcellular pools in the compartmentalized eukaryotic cell. The detection and relative quantification of these individual pools is difficult because of the methods usually applied, which require cell disruption and fractionation.Here, we describe an immunochemical method to visualize and relatively quantify subcellular NAD pools, which relies on the NAD-consuming activity of poly-ADP-ribose polymerase 1 (PARP1). We demonstrate that this system can be readily applied to detect changes in the mitochondrial, Golgi, endoplasmic reticulum, and peroxisomal NAD pools.
烟酰胺腺嘌呤二核苷酸（NAD）对许多细胞过程至关重要，并且在具有区室化结构的真核细胞的不同亚细胞池中分布。由于通常所应用的方法需要细胞破碎和分级分离，因此对这些单个池的检测和相对定量很困难。在此，我们描述了一种免疫化学方法，用于可视化和相对定量亚细胞NAD池，该方法依赖于聚ADP-核糖聚合酶1（PARP1）消耗NAD的活性。我们证明该系统可轻松用于检测线粒体、高尔基体、内质网和过氧化物酶体NAD池的变化。
