On the Weak Binding and Spectroscopic Signature of SARS-CoV-2 nsp14 Interaction with RNA

Author(s)

Hassan, Ayaz ^[1] ; Sedenho, Graziela C. ^[1] ; Vitale, Phelipe A. M. ^[2] ; Oliviera, Mona N. ^[2] ; Crespilho, Frank N. ^[1]

Affiliation

^[1] Univ Sao Paulo, Sao Carlos Inst Chem, Dept Phys Chem, Av Trabalhador Sao Carlense 400, BR-13566590 Sao Carlos, SP - Brazil

^[2] Biolinker, Av Prof Lineu Prestes, BR-05508000 Sao Paulo, SP - Brazil

• Document type: Journal article
• Source: CHEMBIOCHEM; v. 22, n. 24 OCT 2021.
• Web of Science Citations: 0

Abstract

The SARS-CoV-2 non-structural protein 14 (nsp14), known as exoribonuclease is encoded from the large polyprotein of viral genome and is a major constituent of the transcription replication complex (TRC) machinery of the viral RNA synthesis. This protein is highly conserved among the coronaviruses and is a potential target for the development of a therapeutic drug. Here, we report the SARS-CoV-2 nsp14 expression, show its structural characterization, and ss-RNA exonuclease activity through vibrational and electronic spectroscopies. The deconvolution of amide-I band in the FTIR spectrum of the protein revealed a composition of 35 % alpha-helix and 25 % beta-sheets. The binding between protein and RNA is evidenced from the spectral changes in the amide-I region of the nsp14, showing protein conformational changes during the binding process. A value of 20.60 +/- 3.81 mol L-1 of the binding constant (K-D) is obtained for nsp14/RNA complex. The findings reported here can motivate further studies to develop structural models for better understanding the mechanism of exonuclease enzymes for correcting the viral genome and can help in the development of drugs against SARS-CoV-2. (AU)

• FAPESP's process: 20/04796-8 - In situ and operando studies of metaloenzymes for energy conversion and fuel bioelectrosynthesis
• Grantee: Graziela Cristina Sedenho
• Support type: Scholarships in Brazil - Post-Doctorate
• FAPESP's process: 18/22214-6 - Towards a convergence of technologies: from sensing and biosensing to information visualization and machine learning for data analysis in clinical diagnosis
• Grantee: Osvaldo Novais de Oliveira Junior
• Support type: Research Projects - Thematic Grants
• FAPESP's process: 19/12053-8 - High performance electrodes applied in organic batteries and in biofuel cell
• Grantee: Frank Nelson Crespilho
• Support type: Regular Research Grants
• FAPESP's process: 19/15333-1 - Bio-photo-electrochemical hybrid cells for solar energy conversion
• Grantee: Frank Nelson Crespilho
• Support type: Regular Research Grants