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Abstract <p>The proposed approach for determining catalytic activity of the SARS-CoV-2 main protease (M<sup>pro</sup>) is based on registration of the peak area of electrochemical oxidation of tyrosine residue in the model peptide substrate CGGGAVLQSGY immobilized on the surface of a graphite screen-printed electrode (SPE) modified with gold nanoparticles (AuNP). The AuNP were obtained by electrosynthesis. Steady state kinetic parameters of M<sup>pro</sup> in the reaction with the model peptide were determined: catalytic constant&#xa0;(<i>k</i><sub>cat</sub>) was (3.1&#xa0;±&#xa0;0.1)&#xa0;×&#xa0;10<sup>−3</sup>&#xa0;s<sup>−1</sup>; Michaelis constant&#xa0;(<i>K</i><sub>M</sub>) was (358&#xa0;±&#xa0;32)&#xa0;×&#xa0;10<sup>−9</sup>&#xa0;M; catalytic efficiency (<i>k</i><sub>cat</sub>/<i>K</i><sub>M</sub>) was 8659&#xa0;s<sup>−1</sup>/M. The limit of detection (LOD) determined for M<sup>pro</sup> using the proposed electrochemical system was 44&#xa0;nM. The proposed approach is a promising tool to search for new M<sup>pro</sup> inhibitors as drugs for treatment of coronavirus infections.</p>

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Determination of SARS-CoV-2 Main Protease (Mpro) Activity Based on Electrooxidation of Tyrosine Residue of a Model Peptide

  • 期刊论文

  • 发表日期：

    2025年1月1日

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  • Tatiana A. Filippova,
  • Rami A. Masamrekh,
  • Tatiana E. Farafonova,
  • Yulia Yu. Khudoklinova,
  • Victoria V. Shumyantseva,
  • Sergei A. Moshkovskii,
  • Alexey V. Kuzikov

摘要

Abstract

The proposed approach for determining catalytic activity of the SARS-CoV-2 main protease (Mpro) is based on registration of the peak area of electrochemical oxidation of tyrosine residue in the model peptide substrate CGGGAVLQSGY immobilized on the surface of a graphite screen-printed electrode (SPE) modified with gold nanoparticles (AuNP). The AuNP were obtained by electrosynthesis. Steady state kinetic parameters of Mpro in the reaction with the model peptide were determined: catalytic constant (kcat) was (3.1 ± 0.1) × 10−3 s−1; Michaelis constant (KM) was (358 ± 32) × 10−9 M; catalytic efficiency (kcat/KM) was 8659 s−1/M. The limit of detection (LOD) determined for Mpro using the proposed electrochemical system was 44 nM. The proposed approach is a promising tool to search for new Mpro inhibitors as drugs for treatment of coronavirus infections.