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WWOX oxidoreductase--substrate and enzymatic characterization.

Anna Sałuda-Gorgul1, Karolina Seta, Magdalena Nowakowska

  • 1Medical University of Lodz, Department of Analytical Chemistry, Muszynskiego 1, 90-151 Lodz, Poland.

Zeitschrift Fur Naturforschung. C, Journal of Biosciences
|April 12, 2011
PubMed
Summary

The WWOX tumor suppressor gene exhibits oxidoreductase activity, specifically dehydrogenase activity in its SDR domain. This finding sheds light on its potential role in steroid metabolism.

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Published on: October 3, 2018

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • The WWOX gene is a tumor suppressor located at the common fragile site FRA16D.
  • WWOX expression is highest in testis, prostate, and ovary, with altered expression observed in various tumors.
  • The WWOX protein contains a short-chain dehydrogenase/reductase (SDR) domain and WW domains, suggesting a role in steroid metabolism.

Purpose of the Study:

  • To investigate the enzymatic activity of the WWOX protein, particularly its potential role in sex-steroid metabolism.
  • To characterize the oxidoreductase activity of WWOX fusion proteins in vitro.

Main Methods:

  • Cloning of WWOX fusion proteins using two bacterial expression systems.
  • Assessing oxidoreductase activity in crude extracts.
  • Defining enzymatic reaction pathways for steroid substrates.
  • Determining Michaelis-Menten constants (Km) for enzymatic reactions.

Main Results:

  • WWOX fusion proteins demonstrated oxidoreductase activity.
  • The SDR domain of WWOX exhibits significant dehydrogenase activity.
  • This activity was observed with both NAD+ and NADP+ cofactors across all tested steroid substrates.
  • No reduction activity was detected with NADH or NADPH cofactors.

Conclusions:

  • The SDR domain of WWOX possesses specific dehydrogenase activity, crucial for its function.
  • WWOX plays a role in steroid metabolism, likely through its dehydrogenase capabilities.
  • These findings provide biochemical evidence for WWOX's involvement in cellular processes related to steroid transformations.