Phylogenetic and Structure-Function Analyses of ENA ATPases: A Case Study of the ENA1 Protein from the Fungus <i>Neurospora crassa</i>
ENA transporters are a group of P-type ATPases that are characterized by actively moving Na<sup>+</sup> or K<sup>+</sup> out of the cell against their concentration gradient. The existence of these transporters was initially attributed to some fungi, although more recently th...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/1/514 |
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| author | Marcos Aguilella Blanca Garciadeblás Luis Fernández Pacios Begoña Benito |
| author_facet | Marcos Aguilella Blanca Garciadeblás Luis Fernández Pacios Begoña Benito |
| author_sort | Marcos Aguilella |
| collection | DOAJ |
| description | ENA transporters are a group of P-type ATPases that are characterized by actively moving Na<sup>+</sup> or K<sup>+</sup> out of the cell against their concentration gradient. The existence of these transporters was initially attributed to some fungi, although more recently they have also been identified in mosses, liverworts, and some protozoa. Given the current increase in the number of organisms whose genomes are completely sequenced, we set out to expand our knowledge about the existence of ENA in organisms belonging to other phylogenetic groups. For that, a hidden Markov model profile was constructed to identify homologous sequences to ENA proteins in protein databases. This analysis allowed us to identify the existence of ENA-type ATPases in the most primitive groups of fungi, as well as in other eukaryotic organisms not described so far. In addition, this study has allowed the identification of a possible new group of P-ATPases, initially proposed as ENA but which maintain phylogenetic distances with these proteins. Finally, this work has also addressed this study of the structure of ENA proteins, which remained unknown due to the lack of crystallographic data. For this purpose, a 3D structure prediction of the NcENA1 protein of the fungus <i>Neurospora crassa</i> was performed using AlphaFold2 software v2.3.1. From this structure, the electrostatic potential of the protein was analyzed. With all these data, the protein regions and the amino acids involved in the transport of Na<sup>+</sup> or K<sup>+</sup> ions across the membrane were proposed for the first time. Targeted mutagenesis of some of these residues has confirmed their relevant participation in the transport function of ENA proteins. |
| first_indexed | 2024-03-08T15:05:36Z |
| format | Article |
| id | doaj.art-b07c806a308948b59ca89fa4d0c2e77d |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-08T15:05:36Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-b07c806a308948b59ca89fa4d0c2e77d2024-01-10T14:59:48ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-12-0125151410.3390/ijms25010514Phylogenetic and Structure-Function Analyses of ENA ATPases: A Case Study of the ENA1 Protein from the Fungus <i>Neurospora crassa</i>Marcos Aguilella0Blanca Garciadeblás1Luis Fernández Pacios2Begoña Benito3Centro de Biotecnología y Genómica de Plantas (CBGP), Universidad Politécnica de Madrid (UPM), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Campus Montegancedo UPM, Pozuelo de Alarcón, 28223 Madrid, SpainDepartamento de Biotecnología-Biología Vegetal, Escuela Técnica Superior de Ingeniería Agronómica, Alimentaria y de Biosistemas, Universidad Politécnica de Madrid (UPM), 28040 Madrid, SpainCentro de Biotecnología y Genómica de Plantas (CBGP), Universidad Politécnica de Madrid (UPM), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Campus Montegancedo UPM, Pozuelo de Alarcón, 28223 Madrid, SpainCentro de Biotecnología y Genómica de Plantas (CBGP), Universidad Politécnica de Madrid (UPM), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Campus Montegancedo UPM, Pozuelo de Alarcón, 28223 Madrid, SpainENA transporters are a group of P-type ATPases that are characterized by actively moving Na<sup>+</sup> or K<sup>+</sup> out of the cell against their concentration gradient. The existence of these transporters was initially attributed to some fungi, although more recently they have also been identified in mosses, liverworts, and some protozoa. Given the current increase in the number of organisms whose genomes are completely sequenced, we set out to expand our knowledge about the existence of ENA in organisms belonging to other phylogenetic groups. For that, a hidden Markov model profile was constructed to identify homologous sequences to ENA proteins in protein databases. This analysis allowed us to identify the existence of ENA-type ATPases in the most primitive groups of fungi, as well as in other eukaryotic organisms not described so far. In addition, this study has allowed the identification of a possible new group of P-ATPases, initially proposed as ENA but which maintain phylogenetic distances with these proteins. Finally, this work has also addressed this study of the structure of ENA proteins, which remained unknown due to the lack of crystallographic data. For this purpose, a 3D structure prediction of the NcENA1 protein of the fungus <i>Neurospora crassa</i> was performed using AlphaFold2 software v2.3.1. From this structure, the electrostatic potential of the protein was analyzed. With all these data, the protein regions and the amino acids involved in the transport of Na<sup>+</sup> or K<sup>+</sup> ions across the membrane were proposed for the first time. Targeted mutagenesis of some of these residues has confirmed their relevant participation in the transport function of ENA proteins.https://www.mdpi.com/1422-0067/25/1/514ENA ATPasesNa<sup>+</sup> transportersAlphafold2<i>Neurospora crassa</i>phylogeny |
| spellingShingle | Marcos Aguilella Blanca Garciadeblás Luis Fernández Pacios Begoña Benito Phylogenetic and Structure-Function Analyses of ENA ATPases: A Case Study of the ENA1 Protein from the Fungus <i>Neurospora crassa</i> International Journal of Molecular Sciences ENA ATPases Na<sup>+</sup> transporters Alphafold2 <i>Neurospora crassa</i> phylogeny |
| title | Phylogenetic and Structure-Function Analyses of ENA ATPases: A Case Study of the ENA1 Protein from the Fungus <i>Neurospora crassa</i> |
| title_full | Phylogenetic and Structure-Function Analyses of ENA ATPases: A Case Study of the ENA1 Protein from the Fungus <i>Neurospora crassa</i> |
| title_fullStr | Phylogenetic and Structure-Function Analyses of ENA ATPases: A Case Study of the ENA1 Protein from the Fungus <i>Neurospora crassa</i> |
| title_full_unstemmed | Phylogenetic and Structure-Function Analyses of ENA ATPases: A Case Study of the ENA1 Protein from the Fungus <i>Neurospora crassa</i> |
| title_short | Phylogenetic and Structure-Function Analyses of ENA ATPases: A Case Study of the ENA1 Protein from the Fungus <i>Neurospora crassa</i> |
| title_sort | phylogenetic and structure function analyses of ena atpases a case study of the ena1 protein from the fungus i neurospora crassa i |
| topic | ENA ATPases Na<sup>+</sup> transporters Alphafold2 <i>Neurospora crassa</i> phylogeny |
| url | https://www.mdpi.com/1422-0067/25/1/514 |
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