Searching for Frataxin Function: Exploring the Analogy with Nqo15, the Frataxin-like Protein of Respiratory Complex I from <i>Thermus thermophilus</i>
Nqo15 is a subunit of respiratory complex I of the bacterium <i>Thermus thermophilus</i>, with strong structural similarity to human frataxin (FXN), a protein involved in the mitochondrial disease Friedreich’s ataxia (FRDA). Recently, we showed that the expression of recombinant Nqo15 ca...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-02-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/25/3/1912 |
| _version_ | 1797318591378358272 |
|---|---|
| author | Davide Doni Eva Cavallari Martin Ezequiel Noguera Hernan Gustavo Gentili Federica Cavion Gustavo Parisi Maria Silvina Fornasari Geppo Sartori Javier Santos Massimo Bellanda Donatella Carbonera Paola Costantini Marco Bortolus |
| author_facet | Davide Doni Eva Cavallari Martin Ezequiel Noguera Hernan Gustavo Gentili Federica Cavion Gustavo Parisi Maria Silvina Fornasari Geppo Sartori Javier Santos Massimo Bellanda Donatella Carbonera Paola Costantini Marco Bortolus |
| author_sort | Davide Doni |
| collection | DOAJ |
| description | Nqo15 is a subunit of respiratory complex I of the bacterium <i>Thermus thermophilus</i>, with strong structural similarity to human frataxin (FXN), a protein involved in the mitochondrial disease Friedreich’s ataxia (FRDA). Recently, we showed that the expression of recombinant Nqo15 can ameliorate the respiratory phenotype of FRDA patients’ cells, and this prompted us to further characterize both the Nqo15 solution’s behavior and its potential functional overlap with FXN, using a combination of in silico and in vitro techniques. We studied the analogy of Nqo15 and FXN by performing extensive database searches based on sequence and structure. Nqo15’s folding and flexibility were investigated by combining nuclear magnetic resonance (NMR), circular dichroism, and coarse-grained molecular dynamics simulations. Nqo15’s iron-binding properties were studied using NMR, fluorescence, and specific assays and its desulfurase activation by biochemical assays. We found that the recombinant Nqo15 isolated from complex I is monomeric, stable, folded in solution, and highly dynamic. Nqo15 does not share the iron-binding properties of FXN or its desulfurase activation function. |
| first_indexed | 2024-03-08T03:54:33Z |
| format | Article |
| id | doaj.art-4d1d93749a454489a61c6224af669283 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-08T03:54:33Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-4d1d93749a454489a61c6224af6692832024-02-09T15:15:07ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-02-01253191210.3390/ijms25031912Searching for Frataxin Function: Exploring the Analogy with Nqo15, the Frataxin-like Protein of Respiratory Complex I from <i>Thermus thermophilus</i>Davide Doni0Eva Cavallari1Martin Ezequiel Noguera2Hernan Gustavo Gentili3Federica Cavion4Gustavo Parisi5Maria Silvina Fornasari6Geppo Sartori7Javier Santos8Massimo Bellanda9Donatella Carbonera10Paola Costantini11Marco Bortolus12Department of Biology, University of Padova, 35121 Padova, ItalyDepartment of Biology, University of Padova, 35121 Padova, ItalyDepartment of Physiology and Molecular and Cellular Biology, Institute of Biosciences, Biotechnology and Translational Biology (iB3), Faculty of Exact and Natural Sciences, University of Buenos Aires, Intendente Güiraldes 2160, Buenos Aires C1428EG, ArgentinaDepartment of Physiology and Molecular and Cellular Biology, Institute of Biosciences, Biotechnology and Translational Biology (iB3), Faculty of Exact and Natural Sciences, University of Buenos Aires, Intendente Güiraldes 2160, Buenos Aires C1428EG, ArgentinaDepartment of Biology, University of Padova, 35121 Padova, ItalyDepartment of Science and Technology, National University of Quilmes, Roque Saenz Peña 352, Bernal B1876BXD, ArgentinaDepartment of Science and Technology, National University of Quilmes, Roque Saenz Peña 352, Bernal B1876BXD, ArgentinaDepartment of Biomedical Sciences, University of Padova, 35121 Padova, ItalyDepartment of Physiology and Molecular and Cellular Biology, Institute of Biosciences, Biotechnology and Translational Biology (iB3), Faculty of Exact and Natural Sciences, University of Buenos Aires, Intendente Güiraldes 2160, Buenos Aires C1428EG, ArgentinaDepartment of Chemical Sciences, University of Padova, 35131 Padova, ItalyDepartment of Chemical Sciences, University of Padova, 35131 Padova, ItalyDepartment of Biology, University of Padova, 35121 Padova, ItalyDepartment of Chemical Sciences, University of Padova, 35131 Padova, ItalyNqo15 is a subunit of respiratory complex I of the bacterium <i>Thermus thermophilus</i>, with strong structural similarity to human frataxin (FXN), a protein involved in the mitochondrial disease Friedreich’s ataxia (FRDA). Recently, we showed that the expression of recombinant Nqo15 can ameliorate the respiratory phenotype of FRDA patients’ cells, and this prompted us to further characterize both the Nqo15 solution’s behavior and its potential functional overlap with FXN, using a combination of in silico and in vitro techniques. We studied the analogy of Nqo15 and FXN by performing extensive database searches based on sequence and structure. Nqo15’s folding and flexibility were investigated by combining nuclear magnetic resonance (NMR), circular dichroism, and coarse-grained molecular dynamics simulations. Nqo15’s iron-binding properties were studied using NMR, fluorescence, and specific assays and its desulfurase activation by biochemical assays. We found that the recombinant Nqo15 isolated from complex I is monomeric, stable, folded in solution, and highly dynamic. Nqo15 does not share the iron-binding properties of FXN or its desulfurase activation function.https://www.mdpi.com/1422-0067/25/3/1912Friedreich’s ataxiaFrataxinNqo15 |
| spellingShingle | Davide Doni Eva Cavallari Martin Ezequiel Noguera Hernan Gustavo Gentili Federica Cavion Gustavo Parisi Maria Silvina Fornasari Geppo Sartori Javier Santos Massimo Bellanda Donatella Carbonera Paola Costantini Marco Bortolus Searching for Frataxin Function: Exploring the Analogy with Nqo15, the Frataxin-like Protein of Respiratory Complex I from <i>Thermus thermophilus</i> International Journal of Molecular Sciences Friedreich’s ataxia Frataxin Nqo15 |
| title | Searching for Frataxin Function: Exploring the Analogy with Nqo15, the Frataxin-like Protein of Respiratory Complex I from <i>Thermus thermophilus</i> |
| title_full | Searching for Frataxin Function: Exploring the Analogy with Nqo15, the Frataxin-like Protein of Respiratory Complex I from <i>Thermus thermophilus</i> |
| title_fullStr | Searching for Frataxin Function: Exploring the Analogy with Nqo15, the Frataxin-like Protein of Respiratory Complex I from <i>Thermus thermophilus</i> |
| title_full_unstemmed | Searching for Frataxin Function: Exploring the Analogy with Nqo15, the Frataxin-like Protein of Respiratory Complex I from <i>Thermus thermophilus</i> |
| title_short | Searching for Frataxin Function: Exploring the Analogy with Nqo15, the Frataxin-like Protein of Respiratory Complex I from <i>Thermus thermophilus</i> |
| title_sort | searching for frataxin function exploring the analogy with nqo15 the frataxin like protein of respiratory complex i from i thermus thermophilus i |
| topic | Friedreich’s ataxia Frataxin Nqo15 |
| url | https://www.mdpi.com/1422-0067/25/3/1912 |
| work_keys_str_mv | AT davidedoni searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT evacavallari searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT martinezequielnoguera searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT hernangustavogentili searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT federicacavion searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT gustavoparisi searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT mariasilvinafornasari searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT gepposartori searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT javiersantos searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT massimobellanda searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT donatellacarbonera searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT paolacostantini searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi AT marcobortolus searchingforfrataxinfunctionexploringtheanalogywithnqo15thefrataxinlikeproteinofrespiratorycomplexifromithermusthermophilusi |