Generation of Lasso Peptide-Based ClpP Binders
The Clp protease system fulfills a plethora of important functions in bacteria. It consists of a tetradecameric ClpP barrel holding the proteolytic centers and two hexameric Clp-ATPase rings, which recognize, unfold, and then feed substrate proteins into the ClpP barrel for proteolytic degradation....
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/1/465 |
| _version_ | 1797498740662075392 |
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| author | Imran T. Malik Julian D. Hegemann Heike Brötz-Oesterhelt |
| author_facet | Imran T. Malik Julian D. Hegemann Heike Brötz-Oesterhelt |
| author_sort | Imran T. Malik |
| collection | DOAJ |
| description | The Clp protease system fulfills a plethora of important functions in bacteria. It consists of a tetradecameric ClpP barrel holding the proteolytic centers and two hexameric Clp-ATPase rings, which recognize, unfold, and then feed substrate proteins into the ClpP barrel for proteolytic degradation. Flexible loops carrying conserved tripeptide motifs protrude from the Clp-ATPases and bind into hydrophobic pockets (H-pockets) on ClpP. Here, we set out to engineer microcin J25 (MccJ25), a ribosomally synthesized and post-translationally modified peptide (RiPP) of the lasso peptide subfamily, by introducing the conserved tripeptide motifs into the lasso peptide loop region to mimic the Clp-ATPase loops. We studied the capacity of the resulting lasso peptide variants to bind to ClpP and affect its activity. From the nine variants generated, one in particular (12IGF) was able to activate ClpP from <i>Staphylococcus aureus</i> and <i>Bacillus subtilis</i>. While 12IGF conferred stability to ClpP tetradecamers and stimulated peptide degradation, it did not trigger unregulated protein degradation, in contrast to the H-pocket-binding acyldepsipeptide antibiotics (ADEPs). Interestingly, synergistic interactions between 12IGF and ADEP were observed. |
| first_indexed | 2024-03-10T03:37:40Z |
| format | Article |
| id | doaj.art-7d01c89a6dec44a7be4f937456205416 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T03:37:40Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-7d01c89a6dec44a7be4f9374562054162023-11-23T11:40:24ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-12-0123146510.3390/ijms23010465Generation of Lasso Peptide-Based ClpP BindersImran T. Malik0Julian D. Hegemann1Heike Brötz-Oesterhelt2Department of Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, 72076 Tübingen, GermanyHelmholtz Centre for Infection Research (HZI), Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Saarland University Campus, 66123 Saarbrücken, GermanyDepartment of Microbial Bioactive Compounds, Interfaculty Institute of Microbiology and Infection Medicine, University of Tübingen, 72076 Tübingen, GermanyThe Clp protease system fulfills a plethora of important functions in bacteria. It consists of a tetradecameric ClpP barrel holding the proteolytic centers and two hexameric Clp-ATPase rings, which recognize, unfold, and then feed substrate proteins into the ClpP barrel for proteolytic degradation. Flexible loops carrying conserved tripeptide motifs protrude from the Clp-ATPases and bind into hydrophobic pockets (H-pockets) on ClpP. Here, we set out to engineer microcin J25 (MccJ25), a ribosomally synthesized and post-translationally modified peptide (RiPP) of the lasso peptide subfamily, by introducing the conserved tripeptide motifs into the lasso peptide loop region to mimic the Clp-ATPase loops. We studied the capacity of the resulting lasso peptide variants to bind to ClpP and affect its activity. From the nine variants generated, one in particular (12IGF) was able to activate ClpP from <i>Staphylococcus aureus</i> and <i>Bacillus subtilis</i>. While 12IGF conferred stability to ClpP tetradecamers and stimulated peptide degradation, it did not trigger unregulated protein degradation, in contrast to the H-pocket-binding acyldepsipeptide antibiotics (ADEPs). Interestingly, synergistic interactions between 12IGF and ADEP were observed.https://www.mdpi.com/1422-0067/23/1/465Clp proteaseClp ATPaselasso peptideepitope graftingmutagenesisbioengineering |
| spellingShingle | Imran T. Malik Julian D. Hegemann Heike Brötz-Oesterhelt Generation of Lasso Peptide-Based ClpP Binders International Journal of Molecular Sciences Clp protease Clp ATPase lasso peptide epitope grafting mutagenesis bioengineering |
| title | Generation of Lasso Peptide-Based ClpP Binders |
| title_full | Generation of Lasso Peptide-Based ClpP Binders |
| title_fullStr | Generation of Lasso Peptide-Based ClpP Binders |
| title_full_unstemmed | Generation of Lasso Peptide-Based ClpP Binders |
| title_short | Generation of Lasso Peptide-Based ClpP Binders |
| title_sort | generation of lasso peptide based clpp binders |
| topic | Clp protease Clp ATPase lasso peptide epitope grafting mutagenesis bioengineering |
| url | https://www.mdpi.com/1422-0067/23/1/465 |
| work_keys_str_mv | AT imrantmalik generationoflassopeptidebasedclppbinders AT juliandhegemann generationoflassopeptidebasedclppbinders AT heikebrotzoesterhelt generationoflassopeptidebasedclppbinders |