Compartment-Specific Poly-ADP-Ribose Formation as a Biosensor for Subcellular NAD Pools

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.

Keywords: Biosensor; Compartmentalization; Immunoblot analysis; Immunocytochemistry; Mitochondria; NAD; PARP1; Poly-ADP-ribose.