Effect of the drug cyclophosphamide on the activity of porcine kidney betaine aldehyde dehydrogenase

Ramses Cruz-Valencia, Aldo A. Arvizu-Flores, Jesús A. Rosas-Rodríguez, Elisa M. Valenzuela-Soto*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

The enzyme betaine aldehyde dehydrogenase (BADH EC 1.2.1.8) catalyzes the synthesis of glycine betaine (GB), an osmolyte and osmoprotectant. Also, it participates in several metabolic pathways in humans. All BADHs known have cysteine in the active site involved in the aldehyde binding, whereas the porcine kidney enzyme (pkBADH) also has a neighborhood cysteine, both sensitive to oxidation. The antineoplastic and immuno-suppressant pre-drug cyclophosphamide (CTX), and its bioactivation products, have two highly oxidating chlorine atoms. This work aimed to analyze the effect of CTX in the activity of porcine kidney betaine aldehyde dehydrogenase. PkBADH was incubated with varying CTX concentration (0 to 2.0 mM) at 25 °C and lost 50 % of its activity with 2.0 mM CTX. The presence of the coenzyme NAD+ (0.5 mM) decreased 95% the activity in 2.0 mM CTX. The substrate betaine aldehyde (0.05 and 0.4 mM, and the products NADH (0.1–0.5 mM) and GB (1 and 10 mM) did not have an effect on the enzyme inactivation by CTX. The reducing agents, dithiothreitol and β-mercaptoethanol, reverted the pkBADH inactivation, but reduced glutathione (GSH) was unable to restore the enzyme activity. Molecular docking showed that CTX could enter at the enzyme active site, where its chlorine atoms may interact with the catalytic and the neighboring cysteines. The results obtained show that CTX inactivates the pkBADH due to oxidation of the catalytic cysteine or because it oxidizes catalytic and neighborhood cysteine, forming a disulfide bridge with a concomitant decrease in the activity of the enzyme.

Original languageEnglish
Pages (from-to)1467-1475
Number of pages9
JournalMolecular and Cellular Biochemistry
Volume476
Issue number3
DOIs
StateAccepted/In press - 2021

Bibliographical note

Funding Information:
R. Cruz-Valencia gratefully acknowledges a scholarship from CONACyT for graduate studies.

Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer Science+Business Media, LLC part of Springer Nature.

Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.

Keywords

  • Betaine aldehyde dehydrogenase
  • Cyclophosphamide
  • Glycine betaine
  • Inactivation
  • Sulfhydryl oxidation

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