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Innovative Electrochemical Screening Allows Transketolase Inhibitors to Be Identified.

Chloé M G Aymard1, Matilte Halma2, Arnaud Comte1

  • 1Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, ICBMS UMR 5246 CNRS, Université de Lyon, Université Lyon 1, CNRS, INSA Lyon, CPE Lyon, 43 bd du 11 Novembre 1918 , 69622 Villeurbanne Cedex , France.

Analytical Chemistry
|June 29, 2018
PubMed
Summary

A novel electrochemical assay enables rapid, high-throughput screening of transketolase (TK) inhibitors. This assay identified a lead compound inhibiting E. coli TK, offering a promising tool for drug discovery in diseases like cancer and malaria.

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Area of Science:

  • Biochemistry
  • Enzymology
  • Electrochemistry

Background:

  • Transketolases (TKs) are crucial enzymes in the pentose phosphate pathway.
  • TKs are validated therapeutic targets for various diseases, including cancer, tuberculosis, and malaria.
  • Existing TK assays are often slow, enzyme-intensive, and lack specificity.

Purpose of the Study:

  • To develop a novel, high-throughput electrochemical assay for TK activity.
  • To screen a chemical library for TK inhibitors.
  • To characterize the inhibition mechanism of a lead compound.

Main Methods:

  • Development of an electrochemical assay based on oxidative trapping of the TK-thiamine pyrophosphate (TPP) intermediate.
  • Optimization of electrode characteristics, enzyme loading, electrochemical protocol, and substrate concentration.
  • High-throughput screening of 1360 compounds and mechanistic studies.

Main Results:

  • The developed assay allows for 96 parallel measurements in just 7 minutes.
  • A lead compound was identified with an IC50 of 63 μM and a Ki of 3.4 μM against E. coli TK.
  • The assay facilitated the proposal of a TK inhibition mechanism.

Conclusions:

  • The novel electrochemical assay is a rapid and efficient tool for high-throughput screening of TK inhibitors.
  • This assay accelerates the identification of potential drug candidates targeting TK.
  • The findings provide a new avenue for developing therapeutics against TK-related diseases.