Carbanicotinamide adenine dinucleotide: synthesis and enzymological properties of a carbocyclic analog of oxidized nicotinamide adenine dinucleotide

Abstract

The dinucleotide carbanicotinamide adenine dinucleotide (carba-NAD), in which a 2,3-dihydroxycyclopentane ring replaces the beta-D-ribonucleotide ring of the nicotinamide ribonucleoside moiety of NAD, has been synthesized and characterized enzymologically. The synthesis begins with the known 1-aminoribose analogue (+/-)-4 beta-amino-2 alpha,3 alpha-dihydroxy-1 beta-cyclopentanemethanol. The pyridinium ring is first introduced and the resultant nucleoside analogue specifically 5'-phosphorylated. Coupling the racemic carbanicotinamide 5'-mononucleotide with adenosine 5'-monophosphate produces two diastereomeric carba-NAD analogues which are chromatographically separable. Only one diastereomer is a substrate for alcohol dehydrogenase and on this basis is assigned a configuration analogous to D-ribose. The reduced dinucleotide carba-NADH was characterized by fluorescence spectroscopy and found to adopt a "stacked" conformation similar to that of NADH. The analogue is reduced by both yeast and horse liver alcohol dehydrogenase with Km and Vmax values for the analogue close to those observed for NAD. Carba-NAD is resistant to cleavage by NAD glycohydrolase, and the analogue has been demonstrated to noncovalently inhibit the soluble NAD glycohydrolase from Bungarus fasciatus venom at low concentrations (less than or equal to 100 microM).

Role of NAD+ in the Deacetylase Activity of the SIR2-like Proteins

“…Additional evidence for this came from studies with two nonhydrolyzable NAD ϩ analogs. These compounds, carbanicotinamide adenine dinucleotide (carba-NAD ϩ ), and pseudocarba-NAD ϩ (a diastereomer of carba-NAD ϩ ), have a 2,3-di-hydroxycyclopentane ring replacing the ␤-D-ribonucleotide ring of the nicotinamide ribonucleoside moiety of NAD ϩ (14). This results in a nonhydrolyzable pyridinium-N-glycosidic bond between nicotinamide and ADP-ribose that is known to inhibit NAD ϩ glycohydrolases and ADP-ribosyl transferases (15).…”

“…terial DNA ligase (18,19 (14). If simple binding of NAD ϩ to the active site were necessary for deacetylase activity one would expect that the nonhydrolyzable analogs would not be inhibitors of the deacetylation reaction.…”

Studies on CD38 Inhibitors and Their Application to cADPR-Mediated Ca<SUP>2</SUP><SUP>+</SUP> Signaling

“…5) (Slama and Simmons, 1988). It could inhibit the NAD + glycohydrolase activity of Bungarus fasciatus with an IC50 value around 100 M. The replacement of the ring oxygen with carbon in the nicotinamide ribose made the original unstable glycoside bond of nicotinamide more stable and less likely to be hydrolyzed.…”

Fluorescence Properties of Carba Nicotinamide Adenine Dinucleotide for Glucose Sensing

“…As a result cNAD is more stable concerning hydrolytic cleavage than NAD. [12,13] It is well known that cNAD works as coenzyme for enzymatic reactions with glucose dehydrogenase (GlucDH) too. Furthermore, the data given in refs.…”