Abstract
The identity of the product obtained by sodium borohydride reduction of NAD+ as 1,6-dihydro-NAD has been verified by13C NMR spectroscopy. The behaviour of this compound during chromatography on Sephadex G-15 has been found identical with that of the humidity induced inhibitor detected in NADH preparations and postulated to be identical with 1,6-dihydro-NAD. Evidence is presented that this inhibitor is responsible for the bulk of the inhibitory effect developed in moist NADH preparations and that it is generated through a bimolecular reaction between NADH and NAD+ which in essence is a NAD+ catalyzed double bond rearrangement of NADH effected by transfer of hydride anions from NADH to NAD+. One implication of these findings, that removal of NAD+ from NADH preparations should convey increased stability to the reduced cofactor, has been verified. The structure of the lactate dehydrogenase inhibitors described in the literature is discussed.
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Abbreviations
- NADH:
-
1,4-dihydro-nicotinamide adenine dinucleotide
- LDH:
-
lactate dehydrogenase
- ADPR:
-
adenosine 5′-diphosphoribose
- NAD+ :
-
nicotinamide adenine dinucleotide
- 1,6-dihydro-NAD:
-
1,6-dihydronicotinamide adenine dinucleotide
- TMS:
-
tetramethyl silane
- compound 2:
-
4-phosphoryloxy-1,4-dihydro-nicotinamide adenine dinucleotide
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Godtfredsen, S.E., Ottesen, M. & Andersen, N.R. On the mode of formation of 1,6-dihydro-NAD in NADH preparations. Carlsberg Res. Commun. 44, 65 (1979). https://doi.org/10.1007/BF02906522
DOI: https://doi.org/10.1007/BF02906522