A Proteomics-MM/PBSA Dual Approach for the Analysis of SARS-CoV-2 Main Protease Substrate Peptide Specificity

Gloria Gallo, Uilla Barcick, Camila Coelho, Murilo Salardani, Mauricio F. Camacho, Daniela Cajado-Carvalho, Flavio V. Loures, Solange M.T. Serrano, Leon Hardy, Andre Zelanis, Martin Wurtele

Research output: Contribution to journalArticlepeer-review

Abstract

The main protease Mpro of SARS-CoV-2 is a well-studied major drug target. Additionally, it has been linked to this virus’ pathogenicity, possibly through off-target effects. It is also an interesting diagnostic target. To obtain more data on possible substrates as well as to assess the enzyme’s primary specificity a two-step approach was introduced. First, Terminal Amine Isobaric Labeling of Substrates (TAILS) was employed to identify novel Mpro cleavage sites in a mouse lung proteome library. In a second step, using a structural homology model, the MM/PBSA variant MM/GBSA (Molecular Mechanics Poisson-Boltzmann/Generalized Born Surface Area) free binding energy calculations were carried out to determine relevant interacting amino acids. As a result, 58 unique cleavage sites were detected, including six that displayed glutamine at the P1 position. Furthermore, modeling results indicated that Mpro has a far higher potential promiscuity towards substrates than expected. The combination of proteomics and MM/PBSA modeling analysis can thus be useful for elucidating the specificity of Mpro, and thus open novel perspectives for the development of future peptidomimetic drugs against COVID-19, as well as diagnostic tools.

Original languageAmerican English
JournalDefault journal
StatePublished - Jan 1 2022

Keywords

  • Proteomics, TAILS, Free energy calculations, MM/PBSA, COVID-19, Main protease

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