Highly selective tungstate transporter protein TupA from Desulfovibrio alaskensis G20
Abstract Molybdenum and tungsten are taken up by bacteria and archaea as their soluble oxyanions through high affinity transport systems belonging to the ATP-binding cassette (ABC) transporters. The component A (ModA/TupA) of these transporters is the first selection gate from which the cell differe...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2017-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-017-06133-y |
| _version_ | 1818993907313147904 |
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| author | Ana Rita Otrelo-Cardoso Rashmi R. Nair Márcia A. S. Correia Raquel S. Correia Cordeiro Alejandro Panjkovich Dmitri I. Svergun Teresa Santos-Silva Maria G. Rivas |
| author_facet | Ana Rita Otrelo-Cardoso Rashmi R. Nair Márcia A. S. Correia Raquel S. Correia Cordeiro Alejandro Panjkovich Dmitri I. Svergun Teresa Santos-Silva Maria G. Rivas |
| author_sort | Ana Rita Otrelo-Cardoso |
| collection | DOAJ |
| description | Abstract Molybdenum and tungsten are taken up by bacteria and archaea as their soluble oxyanions through high affinity transport systems belonging to the ATP-binding cassette (ABC) transporters. The component A (ModA/TupA) of these transporters is the first selection gate from which the cell differentiates between MoO4 2−, WO4 2− and other similar oxyanions. We report the biochemical characterization and the crystal structure of the apo-TupA from Desulfovibrio desulfuricans G20, at 1.4 Å resolution. Small Angle X-ray Scattering data suggests that the protein adopts a closed and more stable conformation upon ion binding. The role of the arginine 118 in the selectivity of the oxyanion was also investigated and three mutants were constructed: R118K, R118E and R118Q. Isothermal titration calorimetry clearly shows the relevance of this residue for metal discrimination and oxyanion binding. In this sense, the three variants lost the ability to coordinate molybdate and the R118K mutant keeps an extremely high affinity for tungstate. These results contribute to an understanding of the metal-protein interaction, making it a suitable candidate for a recognition element of a biosensor for tungsten detection. |
| first_indexed | 2024-12-20T20:49:31Z |
| format | Article |
| id | doaj.art-ad4b6ab3f0d04375a038f7e830372ad1 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-20T20:49:31Z |
| publishDate | 2017-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-ad4b6ab3f0d04375a038f7e830372ad12022-12-21T19:26:58ZengNature PortfolioScientific Reports2045-23222017-07-017111210.1038/s41598-017-06133-yHighly selective tungstate transporter protein TupA from Desulfovibrio alaskensis G20Ana Rita Otrelo-Cardoso0Rashmi R. Nair1Márcia A. S. Correia2Raquel S. Correia Cordeiro3Alejandro Panjkovich4Dmitri I. Svergun5Teresa Santos-Silva6Maria G. Rivas7UCIBIO/REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de LisboaUCIBIO/REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de LisboaUCIBIO/REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de LisboaUCIBIO/REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de LisboaEuropean Molecular Biology Laboratory-Hamburg Outstation, c/o DESYEuropean Molecular Biology Laboratory-Hamburg Outstation, c/o DESYUCIBIO/REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de LisboaDepartment of Physics, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del LitoralAbstract Molybdenum and tungsten are taken up by bacteria and archaea as their soluble oxyanions through high affinity transport systems belonging to the ATP-binding cassette (ABC) transporters. The component A (ModA/TupA) of these transporters is the first selection gate from which the cell differentiates between MoO4 2−, WO4 2− and other similar oxyanions. We report the biochemical characterization and the crystal structure of the apo-TupA from Desulfovibrio desulfuricans G20, at 1.4 Å resolution. Small Angle X-ray Scattering data suggests that the protein adopts a closed and more stable conformation upon ion binding. The role of the arginine 118 in the selectivity of the oxyanion was also investigated and three mutants were constructed: R118K, R118E and R118Q. Isothermal titration calorimetry clearly shows the relevance of this residue for metal discrimination and oxyanion binding. In this sense, the three variants lost the ability to coordinate molybdate and the R118K mutant keeps an extremely high affinity for tungstate. These results contribute to an understanding of the metal-protein interaction, making it a suitable candidate for a recognition element of a biosensor for tungsten detection.https://doi.org/10.1038/s41598-017-06133-y |
| spellingShingle | Ana Rita Otrelo-Cardoso Rashmi R. Nair Márcia A. S. Correia Raquel S. Correia Cordeiro Alejandro Panjkovich Dmitri I. Svergun Teresa Santos-Silva Maria G. Rivas Highly selective tungstate transporter protein TupA from Desulfovibrio alaskensis G20 Scientific Reports |
| title | Highly selective tungstate transporter protein TupA from Desulfovibrio alaskensis G20 |
| title_full | Highly selective tungstate transporter protein TupA from Desulfovibrio alaskensis G20 |
| title_fullStr | Highly selective tungstate transporter protein TupA from Desulfovibrio alaskensis G20 |
| title_full_unstemmed | Highly selective tungstate transporter protein TupA from Desulfovibrio alaskensis G20 |
| title_short | Highly selective tungstate transporter protein TupA from Desulfovibrio alaskensis G20 |
| title_sort | highly selective tungstate transporter protein tupa from desulfovibrio alaskensis g20 |
| url | https://doi.org/10.1038/s41598-017-06133-y |
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