An engineered protein-based submicromolar competitive inhibitor of the Staphylococcus aureus virulence factor aureolysin
Aureolysin, a secreted metallopeptidase (MP) from the thermolysin family, functions as a major virulence factor in Staphylococcus aureus. No specific aureolysin inhibitors have yet been described, making this an important target for the development of novel antimicrobial drugs in times of rampant an...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-01-01
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| Series: | Computational and Structural Biotechnology Journal |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2001037022000022 |
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| author | Soraia R. Mendes Ulrich Eckhard Arturo Rodríguez-Banqueri Tibisay Guevara Peter Czermak Enrique Marcos Andreas Vilcinskas F. Xavier Gomis-Rüth |
| author_facet | Soraia R. Mendes Ulrich Eckhard Arturo Rodríguez-Banqueri Tibisay Guevara Peter Czermak Enrique Marcos Andreas Vilcinskas F. Xavier Gomis-Rüth |
| author_sort | Soraia R. Mendes |
| collection | DOAJ |
| description | Aureolysin, a secreted metallopeptidase (MP) from the thermolysin family, functions as a major virulence factor in Staphylococcus aureus. No specific aureolysin inhibitors have yet been described, making this an important target for the development of novel antimicrobial drugs in times of rampant antibiotic resistance. Although small-molecule inhibitors are currently more common in the clinic, therapeutic proteins and peptides (TPs) are favourable due to their high selectivity, which reduces off-target toxicity and allows dosage tuning. The greater wax moth Galleria mellonella produces a unique defensive protein known as the insect metallopeptidase inhibitor (IMPI), which selectively inhibits some thermolysins from pathogenic bacteria. We determined the ability of IMPI to inhibit aureolysin in vitro and used crystal structures to ascertain its mechanism of action. This revealed that IMPI uses the “standard mechanism”, which has been poorly characterised for MPs in general. Accordingly, we designed a cohort of 12 single and multiple point mutants, the best of which (I57F) inhibited aureolysin with an estimated inhibition constant (Ki) of 346 nM. Given that animals lack thermolysins, our strategy may facilitate the development of safe TPs against staphylococcal infections, including strains resistant to conventional antibiotics. |
| first_indexed | 2024-04-11T05:20:13Z |
| format | Article |
| id | doaj.art-1e0c32ae4f5848d6a20d93f2fff9494e |
| institution | Directory Open Access Journal |
| issn | 2001-0370 |
| language | English |
| last_indexed | 2024-04-11T05:20:13Z |
| publishDate | 2022-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Computational and Structural Biotechnology Journal |
| spelling | doaj.art-1e0c32ae4f5848d6a20d93f2fff9494e2022-12-24T04:51:08ZengElsevierComputational and Structural Biotechnology Journal2001-03702022-01-0120534544An engineered protein-based submicromolar competitive inhibitor of the Staphylococcus aureus virulence factor aureolysinSoraia R. Mendes0Ulrich Eckhard1Arturo Rodríguez-Banqueri2Tibisay Guevara3Peter Czermak4Enrique Marcos5Andreas Vilcinskas6F. Xavier Gomis-Rüth7Proteolysis Laboratory, Department of Structural Biology, Molecular Biology Institute of Barcelona (CSIC), Barcelona Science Park, Baldiri Reixac 15-21, 08028 Barcelona, Catalonia, SpainProteolysis Laboratory, Department of Structural Biology, Molecular Biology Institute of Barcelona (CSIC), Barcelona Science Park, Baldiri Reixac 15-21, 08028 Barcelona, Catalonia, Spain; Corresponding authors at: Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Ohlebergsweg 12, 35392 Giessen, Germany (A. Vilcinskas).Proteolysis Laboratory, Department of Structural Biology, Molecular Biology Institute of Barcelona (CSIC), Barcelona Science Park, Baldiri Reixac 15-21, 08028 Barcelona, Catalonia, SpainProteolysis Laboratory, Department of Structural Biology, Molecular Biology Institute of Barcelona (CSIC), Barcelona Science Park, Baldiri Reixac 15-21, 08028 Barcelona, Catalonia, SpainDepartment of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Ohlebergsweg 12, 35392 Giessen, Germany; Institute for Insect Biotechnology, Justus-Liebig University Giessen, Heinrich-Buff-Ring 26-32, 35392 Giessen, GermanyProtein Design and Modelling, Department of Structural Biology, Molecular Biology Institute of Barcelona (CSIC), Barcelona Science Park, Baldiri Reixac 15-21, 08028 Barcelona, Catalonia, SpainDepartment of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Ohlebergsweg 12, 35392 Giessen, Germany; Institute for Insect Biotechnology, Justus-Liebig University Giessen, Heinrich-Buff-Ring 26-32, 35392 Giessen, Germany; Corresponding authors at: Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Ohlebergsweg 12, 35392 Giessen, Germany (A. Vilcinskas).Proteolysis Laboratory, Department of Structural Biology, Molecular Biology Institute of Barcelona (CSIC), Barcelona Science Park, Baldiri Reixac 15-21, 08028 Barcelona, Catalonia, Spain; Corresponding authors at: Department of Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology, Ohlebergsweg 12, 35392 Giessen, Germany (A. Vilcinskas).Aureolysin, a secreted metallopeptidase (MP) from the thermolysin family, functions as a major virulence factor in Staphylococcus aureus. No specific aureolysin inhibitors have yet been described, making this an important target for the development of novel antimicrobial drugs in times of rampant antibiotic resistance. Although small-molecule inhibitors are currently more common in the clinic, therapeutic proteins and peptides (TPs) are favourable due to their high selectivity, which reduces off-target toxicity and allows dosage tuning. The greater wax moth Galleria mellonella produces a unique defensive protein known as the insect metallopeptidase inhibitor (IMPI), which selectively inhibits some thermolysins from pathogenic bacteria. We determined the ability of IMPI to inhibit aureolysin in vitro and used crystal structures to ascertain its mechanism of action. This revealed that IMPI uses the “standard mechanism”, which has been poorly characterised for MPs in general. Accordingly, we designed a cohort of 12 single and multiple point mutants, the best of which (I57F) inhibited aureolysin with an estimated inhibition constant (Ki) of 346 nM. Given that animals lack thermolysins, our strategy may facilitate the development of safe TPs against staphylococcal infections, including strains resistant to conventional antibiotics.http://www.sciencedirect.com/science/article/pii/S2001037022000022Bacterial infectionMetallopeptidaseProtein inhibitorProtein designTherapeutic proteinCrystal structure |
| spellingShingle | Soraia R. Mendes Ulrich Eckhard Arturo Rodríguez-Banqueri Tibisay Guevara Peter Czermak Enrique Marcos Andreas Vilcinskas F. Xavier Gomis-Rüth An engineered protein-based submicromolar competitive inhibitor of the Staphylococcus aureus virulence factor aureolysin Computational and Structural Biotechnology Journal Bacterial infection Metallopeptidase Protein inhibitor Protein design Therapeutic protein Crystal structure |
| title | An engineered protein-based submicromolar competitive inhibitor of the Staphylococcus aureus virulence factor aureolysin |
| title_full | An engineered protein-based submicromolar competitive inhibitor of the Staphylococcus aureus virulence factor aureolysin |
| title_fullStr | An engineered protein-based submicromolar competitive inhibitor of the Staphylococcus aureus virulence factor aureolysin |
| title_full_unstemmed | An engineered protein-based submicromolar competitive inhibitor of the Staphylococcus aureus virulence factor aureolysin |
| title_short | An engineered protein-based submicromolar competitive inhibitor of the Staphylococcus aureus virulence factor aureolysin |
| title_sort | engineered protein based submicromolar competitive inhibitor of the staphylococcus aureus virulence factor aureolysin |
| topic | Bacterial infection Metallopeptidase Protein inhibitor Protein design Therapeutic protein Crystal structure |
| url | http://www.sciencedirect.com/science/article/pii/S2001037022000022 |
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