An innovative strategy to investigate microbial protein modifications in a reliable fast and sensitive way: A therapy oriented proof of concept based on UV-C irradiation of SARS-CoV-2 spike protein
The characterization of modifications of microbial proteins is of primary importance to dissect pathogen lifecycle mechanisms and could be useful in identifying therapeutic targets. Attempts to solve this issue yielded only partial and non-exhaustive results. We developed a multidisciplinary approac...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-08-01
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| Series: | Pharmacological Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1043661823002189 |
| _version_ | 1828717009479663616 |
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| author | Sergio Strizzi Letizia Bernardo Pasqualina D’Ursi Chiara Urbinati Andrea Bianco Fiona Limanaqi Andrea Manconi Maria Milanesi Alberto Macchi Dario Di Silvestre Adalberto Cavalleri Giovanni Pareschi Marco Rusnati Mario Clerici PierLuigi Mauri Mara Biasin |
| author_facet | Sergio Strizzi Letizia Bernardo Pasqualina D’Ursi Chiara Urbinati Andrea Bianco Fiona Limanaqi Andrea Manconi Maria Milanesi Alberto Macchi Dario Di Silvestre Adalberto Cavalleri Giovanni Pareschi Marco Rusnati Mario Clerici PierLuigi Mauri Mara Biasin |
| author_sort | Sergio Strizzi |
| collection | DOAJ |
| description | The characterization of modifications of microbial proteins is of primary importance to dissect pathogen lifecycle mechanisms and could be useful in identifying therapeutic targets. Attempts to solve this issue yielded only partial and non-exhaustive results. We developed a multidisciplinary approach by coupling in vitro infection assay, mass spectrometry (MS), protein 3D modelling, and surface plasma resonance (SPR). As a proof of concept, the effect of low UV-C (273 nm) irradiation on SARS-CoV-2 spike (S) protein was investigated. Following UV-C exposure, MS analysis identified, among other modifications, the disruption of a disulphide bond within the conserved S2 subunit of S protein. Computational analyses revealed that this bond breakage associates with an allosteric effect resulting in the generation of a closed conformation with a reduced ability to bind the ACE2 receptor. The UV-C-induced reduced affinity of S protein for ACE2 was further confirmed by SPR analyses and in vitro infection assays. This comprehensive approach pinpoints the S2 domain of S protein as a potential therapeutic target to prevent SARS-CoV-2 infection. Notably, this workflow could be used to screen a wide variety of microbial protein domains, resulting in a precise molecular fingerprint and providing new insights to adequately address future epidemics. |
| first_indexed | 2024-03-12T14:21:44Z |
| format | Article |
| id | doaj.art-b8faaf121c8b47499bce90c1bfe39ffc |
| institution | Directory Open Access Journal |
| issn | 1096-1186 |
| language | English |
| last_indexed | 2024-03-12T14:21:44Z |
| publishDate | 2023-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Pharmacological Research |
| spelling | doaj.art-b8faaf121c8b47499bce90c1bfe39ffc2023-08-19T04:31:32ZengElsevierPharmacological Research1096-11862023-08-01194106862An innovative strategy to investigate microbial protein modifications in a reliable fast and sensitive way: A therapy oriented proof of concept based on UV-C irradiation of SARS-CoV-2 spike proteinSergio Strizzi0Letizia Bernardo1Pasqualina D’Ursi2Chiara Urbinati3Andrea Bianco4Fiona Limanaqi5Andrea Manconi6Maria Milanesi7Alberto Macchi8Dario Di Silvestre9Adalberto Cavalleri10Giovanni Pareschi11Marco Rusnati12Mario Clerici13PierLuigi Mauri14Mara Biasin15Department of Biomedical and Clinical Sciences, University of Milan, Via G.B. Grassi, 20122 Milan, ItalyInstitute for Biomedical Technologies, National Research Council (ITB-CNR), 20054 Segrate, MI, ItalyInstitute for Biomedical Technologies, National Research Council (ITB-CNR), 20054 Segrate, MI, ItalyUnit of Macromolecular Interaction Analysis, Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, ItalyItalian National Institute for Astrophysics (INAF) - Brera Astronomical Observatory, Via E. Bianchi, 46, Merate, 23807 Lecco, ItalyDepartment of Biomedical and Clinical Sciences, University of Milan, Via G.B. Grassi, 20122 Milan, Italy; Department of Pathophysiology and Transplantation, University of Milan, Via Francesco Sforza, 20122 Milan, ItalyInstitute for Biomedical Technologies, National Research Council (ITB-CNR), 20054 Segrate, MI, ItalyInstitute for Biomedical Technologies, National Research Council (ITB-CNR), 20054 Segrate, MI, Italy; Unit of Macromolecular Interaction Analysis, Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, ItalyItalian National Institute for Astrophysics (INAF) - Brera Astronomical Observatory, Via E. Bianchi, 46, Merate, 23807 Lecco, ItalyInstitute for Biomedical Technologies, National Research Council (ITB-CNR), 20054 Segrate, MI, ItalyEpidemiology and Prevention Unit, IRCCS Foundation, Istituto Nazionale dei Tumori, Via Giacomo Venezian, 1, 20133 Milan, ItalyItalian National Institute for Astrophysics (INAF) - Brera Astronomical Observatory, Via E. Bianchi, 46, Merate, 23807 Lecco, ItalyUnit of Macromolecular Interaction Analysis, Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, ItalyDepartment of Pathophysiology and Transplantation, University of Milan, Via Francesco Sforza, 20122 Milan, Italy; Don C. Gnocchi Foundation, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Foundation, Via A. Capecelatro 66, 20148 Milan, íItalyInstitute for Biomedical Technologies, National Research Council (ITB-CNR), 20054 Segrate, MI, Italy; Interdisciplinary Research Center ''Health Science'', Sant’Anna School of Advanced Studies, 56127 Pisa, Italy; Corresponding author at: Institute for Biomedical Technologies, National Research Council (ITB-CNR), 20054 Segrate, MI, Italy.Department of Biomedical and Clinical Sciences, University of Milan, Via G.B. Grassi, 20122 Milan, ItalyThe characterization of modifications of microbial proteins is of primary importance to dissect pathogen lifecycle mechanisms and could be useful in identifying therapeutic targets. Attempts to solve this issue yielded only partial and non-exhaustive results. We developed a multidisciplinary approach by coupling in vitro infection assay, mass spectrometry (MS), protein 3D modelling, and surface plasma resonance (SPR). As a proof of concept, the effect of low UV-C (273 nm) irradiation on SARS-CoV-2 spike (S) protein was investigated. Following UV-C exposure, MS analysis identified, among other modifications, the disruption of a disulphide bond within the conserved S2 subunit of S protein. Computational analyses revealed that this bond breakage associates with an allosteric effect resulting in the generation of a closed conformation with a reduced ability to bind the ACE2 receptor. The UV-C-induced reduced affinity of S protein for ACE2 was further confirmed by SPR analyses and in vitro infection assays. This comprehensive approach pinpoints the S2 domain of S protein as a potential therapeutic target to prevent SARS-CoV-2 infection. Notably, this workflow could be used to screen a wide variety of microbial protein domains, resulting in a precise molecular fingerprint and providing new insights to adequately address future epidemics.http://www.sciencedirect.com/science/article/pii/S1043661823002189UV lightSpike proteinProteomicsStructural biology |
| spellingShingle | Sergio Strizzi Letizia Bernardo Pasqualina D’Ursi Chiara Urbinati Andrea Bianco Fiona Limanaqi Andrea Manconi Maria Milanesi Alberto Macchi Dario Di Silvestre Adalberto Cavalleri Giovanni Pareschi Marco Rusnati Mario Clerici PierLuigi Mauri Mara Biasin An innovative strategy to investigate microbial protein modifications in a reliable fast and sensitive way: A therapy oriented proof of concept based on UV-C irradiation of SARS-CoV-2 spike protein Pharmacological Research UV light Spike protein Proteomics Structural biology |
| title | An innovative strategy to investigate microbial protein modifications in a reliable fast and sensitive way: A therapy oriented proof of concept based on UV-C irradiation of SARS-CoV-2 spike protein |
| title_full | An innovative strategy to investigate microbial protein modifications in a reliable fast and sensitive way: A therapy oriented proof of concept based on UV-C irradiation of SARS-CoV-2 spike protein |
| title_fullStr | An innovative strategy to investigate microbial protein modifications in a reliable fast and sensitive way: A therapy oriented proof of concept based on UV-C irradiation of SARS-CoV-2 spike protein |
| title_full_unstemmed | An innovative strategy to investigate microbial protein modifications in a reliable fast and sensitive way: A therapy oriented proof of concept based on UV-C irradiation of SARS-CoV-2 spike protein |
| title_short | An innovative strategy to investigate microbial protein modifications in a reliable fast and sensitive way: A therapy oriented proof of concept based on UV-C irradiation of SARS-CoV-2 spike protein |
| title_sort | innovative strategy to investigate microbial protein modifications in a reliable fast and sensitive way a therapy oriented proof of concept based on uv c irradiation of sars cov 2 spike protein |
| topic | UV light Spike protein Proteomics Structural biology |
| url | http://www.sciencedirect.com/science/article/pii/S1043661823002189 |
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