tetano
Editor, Senior Moderator
Sci Rep
. 2021 Apr 29;11(1):9283.
doi: 10.1038/s41598-021-88630-9.
The dimer-monomer equilibrium of SARS-CoV-2 main protease is affected by small molecule inhibitors
Lucia Silvestrini[SUP] 1 [/SUP], Norhan Belhaj[SUP] 2 [/SUP], Lucia Comez[SUP] 3 [/SUP], Yuri Gerelli[SUP] 2 [/SUP], Antonino Lauria[SUP] 4 [/SUP], Valeria Libera[SUP] 5 [/SUP], Paolo Mariani[SUP] 2 [/SUP], Paola Marzullo[SUP] 4 [/SUP], Maria Grazia Ortore[SUP] 2 [/SUP], Antonio Palumbo Piccionello[SUP] 4 [/SUP], Caterina Petrillo[SUP] 5 [/SUP], Lucrezia Savini[SUP] 1 [/SUP], Alessandro Paciaroni[SUP] 5 [/SUP], Francesco Spinozzi[SUP] 6 [/SUP]
Affiliations
Abstract
The maturation of coronavirus SARS-CoV-2, which is the etiological agent at the origin of the COVID-19 pandemic, requires a main protease M[SUP]pro[/SUP] to cleave the virus-encoded polyproteins. Despite a wealth of experimental information already available, there is wide disagreement about the M[SUP]pro[/SUP] monomer-dimer equilibrium dissociation constant. Since the functional unit of M[SUP]pro[/SUP] is a homodimer, the detailed knowledge of the thermodynamics of this equilibrium is a key piece of information for possible therapeutic intervention, with small molecules interfering with dimerization being potential broad-spectrum antiviral drug leads. In the present study, we exploit Small Angle X-ray Scattering (SAXS) to investigate the structural features of SARS-CoV-2 M[SUP]pro[/SUP] in solution as a function of protein concentration and temperature. A detailed thermodynamic picture of the monomer-dimer equilibrium is derived, together with the temperature-dependent value of the dissociation constant. SAXS is also used to study how the M[SUP]pro[/SUP] dissociation process is affected by small inhibitors selected by virtual screening. We find that these inhibitors affect dimerization and enzymatic activity to a different extent and sometimes in an opposite way, likely due to the different molecular mechanisms underlying the two processes. The M[SUP]pro[/SUP] residues that emerge as key to optimize both dissociation and enzymatic activity inhibition are discussed.
. 2021 Apr 29;11(1):9283.
doi: 10.1038/s41598-021-88630-9.
The dimer-monomer equilibrium of SARS-CoV-2 main protease is affected by small molecule inhibitors
Lucia Silvestrini[SUP] 1 [/SUP], Norhan Belhaj[SUP] 2 [/SUP], Lucia Comez[SUP] 3 [/SUP], Yuri Gerelli[SUP] 2 [/SUP], Antonino Lauria[SUP] 4 [/SUP], Valeria Libera[SUP] 5 [/SUP], Paolo Mariani[SUP] 2 [/SUP], Paola Marzullo[SUP] 4 [/SUP], Maria Grazia Ortore[SUP] 2 [/SUP], Antonio Palumbo Piccionello[SUP] 4 [/SUP], Caterina Petrillo[SUP] 5 [/SUP], Lucrezia Savini[SUP] 1 [/SUP], Alessandro Paciaroni[SUP] 5 [/SUP], Francesco Spinozzi[SUP] 6 [/SUP]
Affiliations
- PMID: 33927258
- DOI: 10.1038/s41598-021-88630-9
Abstract
The maturation of coronavirus SARS-CoV-2, which is the etiological agent at the origin of the COVID-19 pandemic, requires a main protease M[SUP]pro[/SUP] to cleave the virus-encoded polyproteins. Despite a wealth of experimental information already available, there is wide disagreement about the M[SUP]pro[/SUP] monomer-dimer equilibrium dissociation constant. Since the functional unit of M[SUP]pro[/SUP] is a homodimer, the detailed knowledge of the thermodynamics of this equilibrium is a key piece of information for possible therapeutic intervention, with small molecules interfering with dimerization being potential broad-spectrum antiviral drug leads. In the present study, we exploit Small Angle X-ray Scattering (SAXS) to investigate the structural features of SARS-CoV-2 M[SUP]pro[/SUP] in solution as a function of protein concentration and temperature. A detailed thermodynamic picture of the monomer-dimer equilibrium is derived, together with the temperature-dependent value of the dissociation constant. SAXS is also used to study how the M[SUP]pro[/SUP] dissociation process is affected by small inhibitors selected by virtual screening. We find that these inhibitors affect dimerization and enzymatic activity to a different extent and sometimes in an opposite way, likely due to the different molecular mechanisms underlying the two processes. The M[SUP]pro[/SUP] residues that emerge as key to optimize both dissociation and enzymatic activity inhibition are discussed.