E3 Ubiquitin Ligase SPL2 Is a Lanthanide-Binding Protein
The SPL2 protein is an E3 ubiquitin ligase of unknown function. It is one of only three types of E3 ligases found in the outer membrane of plant chloroplasts. In this study, we show that the cytosolic fragment of SPL2 binds lanthanide ions, as evidenced by fluorescence measurements and circular dich...
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MDPI AG
2021-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/11/5712 |
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| author | Michał Tracz Ireneusz Górniak Andrzej Szczepaniak Wojciech Białek |
| author_facet | Michał Tracz Ireneusz Górniak Andrzej Szczepaniak Wojciech Białek |
| author_sort | Michał Tracz |
| collection | DOAJ |
| description | The SPL2 protein is an E3 ubiquitin ligase of unknown function. It is one of only three types of E3 ligases found in the outer membrane of plant chloroplasts. In this study, we show that the cytosolic fragment of SPL2 binds lanthanide ions, as evidenced by fluorescence measurements and circular dichroism spectroscopy. We also report that SPL2 undergoes conformational changes upon binding of both Ca<sup>2+</sup> and La<sup>3+</sup>, as evidenced by its partial unfolding. However, these structural rearrangements do not interfere with SPL2 enzymatic activity, as the protein retains its ability to auto-ubiquitinate in vitro. The possible applications of lanthanide-based probes to identify protein interactions in vivo are also discussed. Taken together, the results of this study reveal that the SPL2 protein contains a lanthanide-binding site, showing for the first time that at least some E3 ubiquitin ligases are also capable of binding lanthanide ions. |
| first_indexed | 2024-03-10T10:59:05Z |
| format | Article |
| id | doaj.art-0248384c10924de888c5506e139010ba |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T10:59:05Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-0248384c10924de888c5506e139010ba2023-11-21T21:37:00ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-05-012211571210.3390/ijms22115712E3 Ubiquitin Ligase SPL2 Is a Lanthanide-Binding ProteinMichał Tracz0Ireneusz Górniak1Andrzej Szczepaniak2Wojciech Białek3Department of Biophysics, Faculty of Biotechnology, University of Wrocław, Joliot-Curie 14a, 50-383 Wrocław, PolandDepartment of Biophysics, Faculty of Biotechnology, University of Wrocław, Joliot-Curie 14a, 50-383 Wrocław, PolandDepartment of Biophysics, Faculty of Biotechnology, University of Wrocław, Joliot-Curie 14a, 50-383 Wrocław, PolandDepartment of Biophysics, Faculty of Biotechnology, University of Wrocław, Joliot-Curie 14a, 50-383 Wrocław, PolandThe SPL2 protein is an E3 ubiquitin ligase of unknown function. It is one of only three types of E3 ligases found in the outer membrane of plant chloroplasts. In this study, we show that the cytosolic fragment of SPL2 binds lanthanide ions, as evidenced by fluorescence measurements and circular dichroism spectroscopy. We also report that SPL2 undergoes conformational changes upon binding of both Ca<sup>2+</sup> and La<sup>3+</sup>, as evidenced by its partial unfolding. However, these structural rearrangements do not interfere with SPL2 enzymatic activity, as the protein retains its ability to auto-ubiquitinate in vitro. The possible applications of lanthanide-based probes to identify protein interactions in vivo are also discussed. Taken together, the results of this study reveal that the SPL2 protein contains a lanthanide-binding site, showing for the first time that at least some E3 ubiquitin ligases are also capable of binding lanthanide ions.https://www.mdpi.com/1422-0067/22/11/5712ubiquitinationchloroplastlanthanidesSPL2 |
| spellingShingle | Michał Tracz Ireneusz Górniak Andrzej Szczepaniak Wojciech Białek E3 Ubiquitin Ligase SPL2 Is a Lanthanide-Binding Protein International Journal of Molecular Sciences ubiquitination chloroplast lanthanides SPL2 |
| title | E3 Ubiquitin Ligase SPL2 Is a Lanthanide-Binding Protein |
| title_full | E3 Ubiquitin Ligase SPL2 Is a Lanthanide-Binding Protein |
| title_fullStr | E3 Ubiquitin Ligase SPL2 Is a Lanthanide-Binding Protein |
| title_full_unstemmed | E3 Ubiquitin Ligase SPL2 Is a Lanthanide-Binding Protein |
| title_short | E3 Ubiquitin Ligase SPL2 Is a Lanthanide-Binding Protein |
| title_sort | e3 ubiquitin ligase spl2 is a lanthanide binding protein |
| topic | ubiquitination chloroplast lanthanides SPL2 |
| url | https://www.mdpi.com/1422-0067/22/11/5712 |
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