Regulation of the Dimerization and Activity of SARS-CoV-2 Main Protease through Reversible Glutathionylation of Cysteine 300

Davis, David A.; Bulut, Haydar; Shrestha, Prabha; Yaparla, Amulya; Jaeger, Hannah K.; Hattori, Shin-ichiro; Wingfield, Paul T.; Mieyal, John J.; Mitsuya, Hiroaki; Yarchoan, Robert

doi:10.1128/mbio.02094-21

Cited by 15 publications

(20 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similarly, the inhibition efficiency of PRT was reduced in the M165A and Q299A mutants compared to WT. The nature of the direct involvement we identified for Q299 in the inhibition of 3CL pro is consistent with the importance of this region for inhibition by glutathionylation at C300 …”

Section: Resultssupporting

confidence: 82%

Dual-Reporter System for Real-Time Monitoring of SARS-CoV-2 Main Protease Activity in Live Cells Enables Identification of an Allosteric Inhibition Path

Bram

Duan

Nilsson-Payant

et al. 2022

ACS Bio Med Chem Au

View full text Add to dashboard Cite

The SARS-CoV-2 pandemic is an ongoing threat to global health, and the continuing emergence of contagious variants highlights the urgent need for additional antiviral therapy to attenuate COVID-19 disease. The SARS-CoV-2 main protease (3CLpro) presents an attractive target for such therapy due to its high sequence conservation and key role in the viral life cycle. In this study, we designed a fluorescent–luminescent cell-based reporter for the detection and quantification of 3CLpro intracellular activity. Employing this platform, we examined the efficiency of known protease inhibitors against 3CLpro and further identified potent inhibitors through high-throughput chemical screening. Computational analysis confirmed a direct interaction of the lead compounds with the protease catalytic site and identified a prototype for efficient allosteric inhibition. These developments address a pressing need for a convenient sensor and specific targets for both virus detection and rapid discovery of potential inhibitors.

show abstract

Section: Resultssupporting

confidence: 82%

Dual-Reporter System for Real-Time Monitoring of SARS-CoV-2 Main Protease Activity in Live Cells Enables Identification of an Allosteric Inhibition Path

Bram

Duan

Nilsson-Payant

et al. 2022

ACS Bio Med Chem Au

View full text Add to dashboard Cite

show abstract

“… 38 , 39 , 50 , 51 While Cys145 is involved in catalysis, Cys300 has a role in the dimerization of M pro , which is necessary for its catalytic activity. 52 …”

Section: Resultsmentioning

confidence: 99%

The Natural Products Withaferin A and Withanone from the Medicinal HerbWithania somniferaAre Covalent Inhibitors of the SARS-CoV-2 Main Protease

et al. 2022

View full text Add to dashboard Cite

The current COVID-19 pandemic caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) created a global health crisis. The ability of vaccines to protect immunocompromised individuals and from emerging new strains are major concerns. Hence antiviral drugs against SARS-CoV-2 are essential. The SARS-CoV-2 main protease M pro is vital for replication and an important target for antivirals. Using CMap analysis and docking studies, withaferin A (wifA) and withanone (win), two natural products from the medicinal herb Withania somnifera (ashwagandha), were identified as promising candidates that can covalently inhibit the viral protease M pro . Cell culture, enzymatic, LC-MS/MS, computational, and equilibrium dialysis based assays were performed. DFT calculations indicated that wifA and win can form stable adducts with thiols. The cytotoxicity of M pro was significantly reduced by wifA and win. Both wifA and win were found to irreversibly inhibit 0.5 μM M pro with IC 50 values of 0.54 and 1.8 μM, respectively. LC-MS/MS analysis revealed covalent adduct formation with wifA at cysteines 145 and 300 of M pro . The natural products wifA and win can irreversibly inhibit the SARS-CoV-2 main protease M pro . Based on the work presented here we propose that both wifA and win have the potential to be safely used as preventative and therapeutic interventions for COVID-19.

show abstract

“…Compounds containing the thiol group interacted with SARS-CoV-2 in vitro, decreasing virus entry into the cell [ 99 ]. GSH interacts and, in certain conditions, decreases the main SARS-CoV-2 protease activity [ 100 ]. Bioinformatic tools predicted that SARS-CoV-2 main protease targets GPX1, an enzyme involved in neutralizing lipid peroxides and hydrogen peroxide in organisms [ 101 ], as well as a glutamate-cysteine ligase, an enzyme involved in GSH synthesis [ 102 ].…”

Section: Antioxidative Enzymes Studied In Covid-19mentioning

confidence: 99%

Molecular Mechanisms Related to Responses to Oxidative Stress and Antioxidative Therapies in COVID-19: A Systematic Review

et al. 2022

View full text Add to dashboard Cite

The coronavirus disease (COVID-19) pandemic is a leading global health and economic challenge. What defines the disease’s progression is not entirely understood, but there are strong indications that oxidative stress and the defense against reactive oxygen species are crucial players. A big influx of immune cells to the site of infection is marked by the increase in reactive oxygen and nitrogen species. Our article aims to highlight the critical role of oxidative stress in the emergence and severity of COVID-19 and, more importantly, to shed light on the underlying molecular and genetic mechanisms. We have reviewed the available literature and clinical trials to extract the relevant genetic variants within the oxidative stress pathway associated with COVID-19 and the anti-oxidative therapies currently evaluated in the clinical trials for COVID-19 treatment, in particular clinical trials on glutathione and N-acetylcysteine.

show abstract

Regulation of the Dimerization and Activity of SARS-CoV-2 Main Protease through Reversible Glutathionylation of Cysteine 300

Abstract: SARS-CoV-2 is responsible for the devastating COVID-19 pandemic. Therefore, it is imperative that we learn as much as we can about the biochemistry of the coronavirus proteins to inform development of therapy.

Cited by 15 publications

References 51 publications

Dual-Reporter System for Real-Time Monitoring of SARS-CoV-2 Main Protease Activity in Live Cells Enables Identification of an Allosteric Inhibition Path

Dual-Reporter System for Real-Time Monitoring of SARS-CoV-2 Main Protease Activity in Live Cells Enables Identification of an Allosteric Inhibition Path

The Natural Products Withaferin A and Withanone from the Medicinal HerbWithania somniferaAre Covalent Inhibitors of the SARS-CoV-2 Main Protease

Molecular Mechanisms Related to Responses to Oxidative Stress and Antioxidative Therapies in COVID-19: A Systematic Review

Contact Info

Product

Resources

About