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Emissive Synthetic Cofactors: An Isomorphic, Isofunctional, and Responsive NAD+ Analogue.


ABSTRACT: The synthesis, photophysics, and biochemical utility of a fluorescent NAD+ analogue based on an isothiazolo[4,3-d]pyrimidine core (NtzAD+) are described. Enzymatic reactions, photophysically monitored in real time, show NtzAD+ and NtzADH to be substrates for yeast alcohol dehydrogenase and lactate dehydrogenase, respectively, with reaction rates comparable to that of the native cofactors. A drop in fluorescence is seen as NtzAD+ is converted to NtzADH, reflecting a complementary photophysical behavior to that of the native NAD+/NADH. NtzAD+ and NtzADH serve as substrates for NADase, which selectively cleaves the nicotinamide's glycosidic bond yielding tzADP-ribose. NtzAD+ also serves as a substrate for ribosyl transferases, including human adenosine ribosyl transferase 5 (ART5) and Cholera toxin subunit A (CTA), which hydrolyze the nicotinamide and transfer tzADP-ribose to an arginine analogue, respectively. These reactions can be monitored by fluorescence spectroscopy, in stark contrast to the corresponding processes with the nonemissive NAD+.

SUBMITTER: Rovira AR 

PROVIDER: S-EPMC5766281 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature

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Emissive Synthetic Cofactors: An Isomorphic, Isofunctional, and Responsive NAD<sup>+</sup> Analogue.

Rovira Alexander R AR   Fin Andrea A   Tor Yitzhak Y  

Journal of the American Chemical Society 20171027 44


The synthesis, photophysics, and biochemical utility of a fluorescent NAD<sup>+</sup> analogue based on an isothiazolo[4,3-d]pyrimidine core (N<sup>tz</sup>AD<sup>+</sup>) are described. Enzymatic reactions, photophysically monitored in real time, show N<sup>tz</sup>AD<sup>+</sup> and N<sup>tz</sup>ADH to be substrates for yeast alcohol dehydrogenase and lactate dehydrogenase, respectively, with reaction rates comparable to that of the native cofactors. A drop in fluorescence is seen as N<sup>tz  ...[more]

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