Redox Coenzymes

No.165(April 2015)

T2979: Thionicotinamide Adenine Dinucleotide oxidized form (1)
T2980: Thionicotinamide Adenine Dinucleotide Disodium Salt reduced form (2)

Enzymatic cycling methods have been designed for amplifying sensitivity to assay a small amount of substrate or enzyme activity.¹⁾ Earlier studies on enzymatic cycling methods were described on DAN⁺, NADP⁺, and CoA.^2-4) Thio-NAD⁺ (1) contributed to improvement of the enzymatic cycling method.^1,5)

Conversion of MVA to HMG-CoA by HMG-CoA reductase⁶⁾

An example of application of 1 for the enzymatic cycling method is shown above. As 1 is converted to thio-NADH (2) by such an enzymatic reaction, the reaction can be traced by the production of 2. Accumulation of 2 can be monitored by the increase in its absorbance (400 nm), which is at a longer wavelength than NADH (340nm).⁸⁾ On the other hand, 2 was also applied for analysis of the active form of bleomycin.⁹⁾

References

1)Carnitine determination by an enzymatic cycling method with carnitine dehydrogenase
- M. Takahashi, S. Ueda, H. Misaki, N. Sugiyama, K. Matsumoto, N. Matsuo, S. Murao, Clin. Chem. 1994, 40, 817.
2)An enzymatic cycling method for nicotinamide-adenine dinucleotide with malic and alcohol dehydrogenases
- T. Kato, S. J. Berger, J. A. Carter, O. H. Lowry, Anal. Biochem. 1973, 53, 86.
3)The measurement of pyridine nucleotides by enzymatic cycling
- O. H. Lowry, J. V. Passonneau, D. W. Schulz, M. K. Rock, J. Biol. Chem. 1961, 236, 2746.
4)Distribution of enzymes between nucleus and cytoplasm of single nerve cell bodies
- T. Kato, O. H. Lowry, J. Biol. Chem. 1973, 248, 2044.
5)ALT reagent with thionicotinamide adenine dinucleotide
- P. A. Dolan, C. D. Pennington, D. A. Yost, Clin. Chem. 1989, 35, 1857.
6)An ultrasensitive enzymatic method for measuring mevalonic acid in serum
- T. Matsuoka, S. Ueda, H. Matsumoto, M. Kawakami, J. Lipid Res. 2012, 53, 1987.
7)Spectrophotometric method for the assay of steroid 5-reductase activity of rat liver and prostate microsome
- A. Iwai, T. Yoshimura, K. Wada, S. Watabe, Y. Sakamoto, E. Ito, T. Miura, Anal. Sci. 2013, 29, 455.
8)Ultrasensitive enzyme-linked immunosorbent assay (ELISA) of proteins by combination with the thio-NAD cycling method
- S. Watabe, H. Kodama, M. Kaneda, M. Morikawa, K. Nakaishi, T. Yoshimura, A. Iwai, T. Miura, E. Ito, Biophysics 2014, 10, 49.
9)The redox state of activated bleomycin
- R. M. Burger, J. S. Blanchard, S. B. Horwitz, J. Peisach, J. Biol. Chem. 1985, 260, 15406.

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Redox Coenzymes

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