Insights into the maturation of the main SARS-CoV-2 protease

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F24%3A00604162" target="_blank" >RIV/61388963:_____/24:00604162 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Insights into the maturation of the main SARS-CoV-2 protease

Popis výsledku v původním jazyce

Mpro, the main protease of SARS-CoV-2, is an important drug target utilized in COVID-19 treatment. Mpro is vital for the effective release of functional proteins, undergoing autocatalytic activation upon release from the viral polyprotein. This process involves dimerization and self-cleavage of the protease. Despite the significance of the enzyme in the viral life cycle, the activation process is still not fully understood. However, targeting the initial step of the proteolytic cascade may reveal new opportunities for COVID-19 treatment. We constructed artificial Mpro precursors to better understand Mpro autoprocessing and investigate the possibilities of Mpro precursor inhibition. To this end, we evaluated the antiviral drugs nirmatrelvir, ensitrelvir, and several in-house designed compounds with the aim of identifying differences in inhibitor susceptibility between the mature and precursor forms of Mpro. nThe Mpro precursor was examined using a bicistronic bacterial expression system followed by Western blot analysis and a reporter cell-based assay. The assay involved transfecting HEK293T cells with a protease fused with two adjacent fluorescent proteins. Finally, we analyzed differences between mature and precursor inhibition using flow cytometry and microscopic techniques. These experiments aim to deepen our understanding of the coronavirus life cycle, offering new insights into drug design.nn

Název v anglickém jazyce

Insights into the maturation of the main SARS-CoV-2 protease

Popis výsledku anglicky

Mpro, the main protease of SARS-CoV-2, is an important drug target utilized in COVID-19 treatment. Mpro is vital for the effective release of functional proteins, undergoing autocatalytic activation upon release from the viral polyprotein. This process involves dimerization and self-cleavage of the protease. Despite the significance of the enzyme in the viral life cycle, the activation process is still not fully understood. However, targeting the initial step of the proteolytic cascade may reveal new opportunities for COVID-19 treatment. We constructed artificial Mpro precursors to better understand Mpro autoprocessing and investigate the possibilities of Mpro precursor inhibition. To this end, we evaluated the antiviral drugs nirmatrelvir, ensitrelvir, and several in-house designed compounds with the aim of identifying differences in inhibitor susceptibility between the mature and precursor forms of Mpro. nThe Mpro precursor was examined using a bicistronic bacterial expression system followed by Western blot analysis and a reporter cell-based assay. The assay involved transfecting HEK293T cells with a protease fused with two adjacent fluorescent proteins. Finally, we analyzed differences between mature and precursor inhibition using flow cytometry and microscopic techniques. These experiments aim to deepen our understanding of the coronavirus life cycle, offering new insights into drug design.nn

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/LX22NPO5103" target="_blank" >LX22NPO5103: Národní institut virologie a bakteriologie</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2024

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů