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

IF 2.3 4区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry (Moscow) Pub Date : 2025-03-07 DOI:10.1134/S0006297924603836

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

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引用次数: 0

Abstract

The proposed approach for determining catalytic activity of the SARS-CoV-2 main protease (M^pro) 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^pro in the reaction with the model peptide were determined: catalytic constant (k_cat) was (3.1 ± 0.1) × 10⁻³ s⁻¹; Michaelis constant (K_M) was (358 ± 32) × 10⁻⁹ M; catalytic efficiency (k_cat/K_M) was 8659 s⁻¹/M. The limit of detection (LOD) determined for M^pro using the proposed electrochemical system was 44 nM. The proposed approach is a promising tool to search for new M^pro inhibitors as drugs for treatment of coronavirus infections.

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来源期刊

Biochemistry (Moscow) 生物-生化与分子生物学

CiteScore

4.70

自引率

3.60%

发文量

139

审稿时长

2 months

期刊介绍： Biochemistry (Moscow) is the journal that includes research papers in all fields of biochemistry as well as biochemical aspects of molecular biology, bioorganic chemistry, microbiology, immunology, physiology, and biomedical sciences. Coverage also extends to new experimental methods in biochemistry, theoretical contributions of biochemical importance, reviews of contemporary biochemical topics, and mini-reviews (News in Biochemistry).