Abstract
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.
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Acknowledgments
MVL, MN, MZ, and CD were supported by the Research Council of Norway (project #. 214435). A.A.N. was supported by the Russian Science Foundation (Grant # 16-14-10240).
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VanLinden, M.R., Niere, M., Nikiforov, A.A., Ziegler, M., Dölle, C. (2017). Compartment-Specific Poly-ADP-Ribose Formation as a Biosensor for Subcellular NAD Pools. In: Tulin, A. (eds) Poly(ADP-Ribose) Polymerase. Methods in Molecular Biology, vol 1608. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6993-7_4
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DOI: https://doi.org/10.1007/978-1-4939-6993-7_4
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-6993-7
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