Although protein ADP-ribosylation is involved in diverse biological processes, it has remained a challenge to identify ADP-ribose acceptor sites. Here, we present an experimental workflow for sensitive and unbiased analysis of endogenous ADP-ribosylation sites, capable of detecting more than 900 modification sites in mammalian cells and mouse liver. In cells, we demonstrate that Lys residues, b...

We have characterized the covalent poly(ADP-ribosyl)ation of p53 using an in vitro reconstituted system. We used recombinant wild type p53, recombinant poly(ADP-ribose) polymerase-1 (PARP-1) (EC ), and betaNAD(+). Our results show that the covalent poly(ADP-ribosyl)ation of p53 is a time-dependent protein-poly(ADP-ribosyl)ation reaction and that the addition of this tumor suppressor protein to ...

Alcoholic beverages like apple cider are considered functional with several health benefits, when consumed in moderation, which mainly attributed to their microbiota and the plethora of bioactive compounds. Among them, bio-functional polar lipids (PL) have recently been found cider, despite low quantities, exhibited strong anti-inflammatory anti-platelet properties, while fermentation seems aff...

Mutants resistant to 3-aminobenzamide, a known inhibitor of ADP-ribosyltransferase, were obtained from Streptomyces coelicolor A3(2). One (strain 27) was analyzed in detail. Mutant 27 had a reduced ADP-ribosyl-transferase activity, exhibited substantial changes from the wild type in ADP-ribosylated protein profile during cell aging, and was defective in producing aerial mycelium and antibiotics...

The accumulation of alpha-1,4-polyglucans is an important strategy to cope with transient starvation conditions in the environment. In bacteria and plants, the synthesis of glycogen and starch occurs by utilizing ADP-glucose as the glucosyl donor for elongation of the alpha-1,4-glucosidic chain. The main regulatory step takes place at the level of ADP-glucose synthesis, a reaction catalyzed by ...

ADP-ribosyltransferases (ARTs) modify proteins with single units or polymers of ADP-ribose to regulate DNA repair. However, the substrates for these enzymes are ill-defined. For example, although histones are modified by ARTs, the sites on these proteins ADP-ribosylated following DNA damage and the ARTs that catalyse these events are unknown. This, in part, is due to the lack of a eukaryotic mo...

Niacin is required to form NAD and NADP, which are involved in many essential redox reactions in cellular metabolism. In addition, NAD acts as a substrate for a variety of ADP-ribosylation reactions, including polyandmono-ADP-ribosylation of proteins, formation of cyclic ADP-ribose, and the generation of O-acetyl-ADP-ribose in deacetylation reactions. These nonredox reactions are critical in th...

Mono-ADP-ribosylation is a reversible posttranslational modification that modulates the function of target proteins. The enzymes that catalyze this reaction in mammalian cells are either bacterial pathogenic toxins or endogenous cellular ADP-ribosyltransferases. For the latter, both the enzymes and their targets have largely remained elusive, mainly due to the lack of specific techniques to stu...

With suitable submitochondrial particles from the heart, the rate of the inorganic phosphate e HOH exchange during substrate oxidation in the absence of ADP is less than l/1000 of the rate in the presence of ADP. The concentration dependence of the exchange on ADP shows simple saturation behavior with an apparent K, of 0.32 mM. For net oxidative phosphorylation under similar conditions, but wit...

With suitable submitochondrial particles from the heart, the rate of the inorganic phosphate e HOH exchange during substrate oxidation in the absence of ADP is less than l/1000 of the rate in the presence of ADP. The concentration dependence of the exchange on ADP shows simple saturation behavior with an apparent K, of 0.32 mM. For net oxidative phosphorylation under similar conditions, but wit...