Journal article

Publication year: 1999

Pages: 1-8

Journal: Analytical Biochemistry

Volume number: 268

Issue number: 1

ISSN: 0003-2697

DOI Link: https://doi.org/10.1006/abio.1998.3061

Publisher: Elsevier

Abstract: 
Trypanothione reductases (TR; EC 1.6.4.8) and thioredoxin reductases (TrxR; EC 1.6.4.5.) are enzymes central to cellular thiol metabolism. Trypanosoma cruzi TR (TcTR) is therefore considered as a potential candidate for drug design against trypanosomiasis. Inhibition of human TrxR (hTrxR) is likely to be beneficial in psoriasis, cancer, and autoimmune diseases, while inhibition of a putative TrxR from Plasmodium falciparum (PfTxR) might prove effective against malaria. The natural substrates of the first two enzymes are very expensive and difficult to obtain; in the case of PfTrxR, the physiological substrate has not yet been identified. We have therefore synthesized and tested three different 5,5'-dithiobis(2-nitrobenzamides) as alternative substrates of the above enzymes. As with 5,5'-dithiobis(2-nitrobenzoate) (DTNB), which can be reduced by TRs and TrxRs, the new compounds are converted to their corresponding chromophoric thiolates; however, they have much lower K-m values and are therefore less likely to interfere with inhibitor testing. Using the new substrates, a novel enzyme assay has been developed which is identical for all three enzymes, can be performed in a microtiter plate, and is amenable to automation. Thus, the assay provides a versatile and inexpensive tool for kinetic studies and high-throughput inhibitor screening. (C) 1999 Academic Press.

Harvard Citation style: Davioud-Charvet, E., Becker, K., Landry, V., Gromer, S., Loge, C. and Sergheraert, C. (1999) Synthesis of 5,5 '-dithiobis(2-nitrobenzamides) as alternative substrates for trypanothione reductase and thioredoxin reductase: A microtiter colorimetric assay for inhibitor screening, Analytical Biochemistry, 268(1), pp. 1-8. https://doi.org/10.1006/abio.1998.3061

APA Citation style: Davioud-Charvet, E., Becker, K., Landry, V., Gromer, S., Loge, C., & Sergheraert, C. (1999). Synthesis of 5,5 '-dithiobis(2-nitrobenzamides) as alternative substrates for trypanothione reductase and thioredoxin reductase: A microtiter colorimetric assay for inhibitor screening. Analytical Biochemistry. 268(1), 1-8. https://doi.org/10.1006/abio.1998.3061