Identification of peptides interfering with the LRRK2/PP1 interaction.
Serine/threonine phosphatases are responsible for modulating the activities of the protein kinases implicated in the development of several pathologies. Here we identified by a PEP-scan approach a peptide of LRRK2, a Parkinson's disease associated protein, interacting with the phosphatase PP1....
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2020-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0237110 |
| _version_ | 1819111646464835584 |
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| author | Chang Zhi Dong Heriberto Bruzzoni-Giovanelli Yanhua Yu Karim Dorgham Christophe Parizot Jean Marc Zini Jean Yves Brossas Pierre Tuffery Angelita Rebollo |
| author_facet | Chang Zhi Dong Heriberto Bruzzoni-Giovanelli Yanhua Yu Karim Dorgham Christophe Parizot Jean Marc Zini Jean Yves Brossas Pierre Tuffery Angelita Rebollo |
| author_sort | Chang Zhi Dong |
| collection | DOAJ |
| description | Serine/threonine phosphatases are responsible for modulating the activities of the protein kinases implicated in the development of several pathologies. Here we identified by a PEP-scan approach a peptide of LRRK2, a Parkinson's disease associated protein, interacting with the phosphatase PP1. In order to study its biological activity, the peptide was fused via its N-terminal to an optimized cell penetrating peptide. We synthesized from the original peptide five interfering peptides and identified two (Mut3DPT-LRRK2-Short and Mut3DPT-LRRK2-Long) able to disrupt the LRRK2/PP1 interaction by competition in anti-LRRK2 immunoprecipitates. Using FITC-labelled peptides, we confirmed their internalization into cell lines as well as into primary cells obtained from healthy or ill human donors. We confirmed by ELISA test the association of Mut3DPT-LRRK2-Long peptide to purified PP1 protein. The peptides Mut3DPT-LRRK2-5 to 8 with either N or C-terminal deletions were not able to disrupt the association LRRK2/PP1 nor to associate with purified PP1 protein. The interfering sequences blocking the PP1/LRRK2 interaction were also fused to a shuttle peptide able to cross the blood brain barrier and showed that the newly generated peptides BBB-LRRK2-Short and BBB-LRRK2-Long were highly resistant to protease degradation. Furthermore, they blocked PP1/LRRK2 interaction and they penetrated into cells. Hence, these newly generated peptides can be employed as new tools in the investigation of the role of the LRRK2/PP1 interaction in normal and pathological conditions. |
| first_indexed | 2024-12-22T04:00:55Z |
| format | Article |
| id | doaj.art-4d97c334c4de49e08f3d32c1061afd4e |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-22T04:00:55Z |
| publishDate | 2020-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-4d97c334c4de49e08f3d32c1061afd4e2022-12-21T18:39:44ZengPublic Library of Science (PLoS)PLoS ONE1932-62032020-01-01158e023711010.1371/journal.pone.0237110Identification of peptides interfering with the LRRK2/PP1 interaction.Chang Zhi DongHeriberto Bruzzoni-GiovanelliYanhua YuKarim DorghamChristophe ParizotJean Marc ZiniJean Yves BrossasPierre TufferyAngelita RebolloSerine/threonine phosphatases are responsible for modulating the activities of the protein kinases implicated in the development of several pathologies. Here we identified by a PEP-scan approach a peptide of LRRK2, a Parkinson's disease associated protein, interacting with the phosphatase PP1. In order to study its biological activity, the peptide was fused via its N-terminal to an optimized cell penetrating peptide. We synthesized from the original peptide five interfering peptides and identified two (Mut3DPT-LRRK2-Short and Mut3DPT-LRRK2-Long) able to disrupt the LRRK2/PP1 interaction by competition in anti-LRRK2 immunoprecipitates. Using FITC-labelled peptides, we confirmed their internalization into cell lines as well as into primary cells obtained from healthy or ill human donors. We confirmed by ELISA test the association of Mut3DPT-LRRK2-Long peptide to purified PP1 protein. The peptides Mut3DPT-LRRK2-5 to 8 with either N or C-terminal deletions were not able to disrupt the association LRRK2/PP1 nor to associate with purified PP1 protein. The interfering sequences blocking the PP1/LRRK2 interaction were also fused to a shuttle peptide able to cross the blood brain barrier and showed that the newly generated peptides BBB-LRRK2-Short and BBB-LRRK2-Long were highly resistant to protease degradation. Furthermore, they blocked PP1/LRRK2 interaction and they penetrated into cells. Hence, these newly generated peptides can be employed as new tools in the investigation of the role of the LRRK2/PP1 interaction in normal and pathological conditions.https://doi.org/10.1371/journal.pone.0237110 |
| spellingShingle | Chang Zhi Dong Heriberto Bruzzoni-Giovanelli Yanhua Yu Karim Dorgham Christophe Parizot Jean Marc Zini Jean Yves Brossas Pierre Tuffery Angelita Rebollo Identification of peptides interfering with the LRRK2/PP1 interaction. PLoS ONE |
| title | Identification of peptides interfering with the LRRK2/PP1 interaction. |
| title_full | Identification of peptides interfering with the LRRK2/PP1 interaction. |
| title_fullStr | Identification of peptides interfering with the LRRK2/PP1 interaction. |
| title_full_unstemmed | Identification of peptides interfering with the LRRK2/PP1 interaction. |
| title_short | Identification of peptides interfering with the LRRK2/PP1 interaction. |
| title_sort | identification of peptides interfering with the lrrk2 pp1 interaction |
| url | https://doi.org/10.1371/journal.pone.0237110 |
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