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Campo DC | Valor | Lengua/Idioma |
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dc.contributor.author | Sanguinetti Miralles, Manuel | - |
dc.contributor.author | Silva Santos, Lucianna Helene | - |
dc.contributor.author | Dourron, Juliette | - |
dc.contributor.author | Alamón Lima, Catalina Isadora | - |
dc.contributor.author | Idiarte Montiel, Juan | - |
dc.contributor.author | Amillis, Sotiris | - |
dc.contributor.author | Pantano, Sergio | - |
dc.contributor.author | Ramón, Ana | - |
dc.date.accessioned | 2023-11-21T12:01:21Z | - |
dc.date.available | 2023-11-21T12:01:21Z | - |
dc.date.issued | 2022 | - |
dc.identifier.citation | Sanguinetti Miralles, M, Silva Santos, L, Dourron, J, [y otros autores]. "Substrate recognition properties from an intermediate structural state of the UreA transporter". International Journal of Molecular Science. [en línea] 2022, 23(24): 16039. 17 h. DOI: 10.3390/ijms232416039 | es |
dc.identifier.issn | 1422-0067 | - |
dc.identifier.uri | https://hdl.handle.net/20.500.12008/41370 | - |
dc.description.abstract | Through a combination of comparative modeling, site-directed and classical random mutagenesis approaches, we previously identified critical residues for binding, recognition, and translocation of urea, and its inhibition by 2-thiourea and acetamide in the Aspergillus nidulans urea transporter, UreA. To deepen the structural characterization of UreA, we employed the artificial intelligence (AI) based AlphaFold2 (AF2) program. In this analysis, the resulting AF2 models lacked inward- and outward-facing cavities, suggesting a structural intermediate state of UreA. Moreover, the orientation of the W82, W84, N279, and T282 side chains showed a large variability, which in the case of W82 and W84, may operate as a gating mechanism in the ligand pathway. To test this hypothesis non-conservative and conservative substitutions of these amino acids were introduced, and binding and transport assessed for urea and its toxic analogue 2-thiourea, as well as binding of the structural analogue acetamide. As a result, residues W82, W84, N279, and T282 were implicated in substrate identification, selection, and translocation. Using molecular docking with Autodock Vina with flexible side chains, we corroborated the AF2 theoretical intermediate model, showing a remarkable correlation between docking scores and experimental affinities determined in wild-type and UreA mutants. The combination of AI-based modeling with classical docking, validated by comprehensive mutational analysis at the binding region, would suggest an unforeseen option to determine structural level details on a challenging family of proteins. | es |
dc.description.sponsorship | ANII: FCE_3_2018_1_148002 | es |
dc.format.extent | 17 h. | es |
dc.format.mimetype | application/pdf | es |
dc.language.iso | en_US | es |
dc.publisher | MDPI | es |
dc.relation.ispartof | International Journal of Molecular Science, 2022, 23(24): 16039. | es |
dc.rights | Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014) | es |
dc.subject | Aspergillus nidulans | es |
dc.subject | Urea transport | es |
dc.subject | AlphaFold2 | es |
dc.subject | Binding site | es |
dc.subject | Molecular docking | es |
dc.title | Substrate recognition properties from an intermediate structural state of the UreA transporter | es |
dc.type | Artículo | es |
dc.contributor.filiacion | Sanguinetti Miralles Manuel, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. | - |
dc.contributor.filiacion | Silva Santos Lucianna Helene, Instituto Pasteur (Montevideo). | - |
dc.contributor.filiacion | Dourron Juliette, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. | - |
dc.contributor.filiacion | Alamón Lima Catalina Isadora, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. | - |
dc.contributor.filiacion | Idiarte Montiel Juan, Instituto Pasteur (Montevideo). | - |
dc.contributor.filiacion | Amillis Sotiris | - |
dc.contributor.filiacion | Pantano Sergio, Instituto Pasteur (Montevideo). | - |
dc.contributor.filiacion | Ramón Ana, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. | - |
dc.rights.licence | Licencia Creative Commons Atribución (CC - By 4.0) | es |
dc.identifier.doi | 10.3390/ijms232416039 | - |
Aparece en las colecciones: | Publicaciones académicas y científicas - Facultad de Ciencias |
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103390ijms232416039.pdf | 2,54 MB | Adobe PDF | Visualizar/Abrir |
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