Key Amino Acid Residues of the Agt1 Transporter for Trehalose Transport by <i>Saccharomyces cerevisiae</i>
Trehalose is crucial for the stress resistance of <i>Saccharomyces cerevisiae</i>, primarily through its stabilization of proteins and membranes. The Agt1 transporter, a member of the Major Facilitator Superfamily, mediates trehalose uptake, a key process for maintaining cellular integri...
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MDPI AG
2024-11-01
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| Series: | Journal of Fungi |
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| Online Access: | https://www.mdpi.com/2309-608X/10/11/781 |
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| author | Anqi Chen Yuhan Cheng Liushi Meng Jian Chen |
| author_facet | Anqi Chen Yuhan Cheng Liushi Meng Jian Chen |
| author_sort | Anqi Chen |
| collection | DOAJ |
| description | Trehalose is crucial for the stress resistance of <i>Saccharomyces cerevisiae</i>, primarily through its stabilization of proteins and membranes. The Agt1 transporter, a member of the Major Facilitator Superfamily, mediates trehalose uptake, a key process for maintaining cellular integrity under stress. Despite its importance, the molecular mechanisms of Agt1-mediated trehalose transport remain underexplored. In this study, we expressed and purified the trehalase enzyme TreA from <i>E. coli</i> to develop reliable trehalose assays. We screened 257 wild <i>S. cerevisiae</i> isolates, identifying strains with enhanced trehalose transport capacities. Comparative analyses, including structural modeling and molecular docking, revealed that specific Agt1 variants exhibited significantly higher transport efficiency, influenced by key residues in the transporter. Molecular dynamics simulations and steered molecular dynamics provided further insights, particularly into the role of the Agt1 channel head region in substrate recognition and binding. Site-directed mutagenesis validated these findings, showing that mutations at critical residues, such as 156Q, 164L, 256Q, 395E, 396R, and 507Y significantly reduced transport activity, while 137Q, 230T, and 514 N increased efficiency under certain conditions. |
| format | Article |
| id | doaj-art-5d21cac739124f7f9f81e70d24a305c9 |
| institution | Kabale University |
| issn | 2309-608X |
| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
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| series | Journal of Fungi |
| spelling | doaj-art-5d21cac739124f7f9f81e70d24a305c92024-11-26T18:08:51ZengMDPI AGJournal of Fungi2309-608X2024-11-01101178110.3390/jof10110781Key Amino Acid Residues of the Agt1 Transporter for Trehalose Transport by <i>Saccharomyces cerevisiae</i>Anqi Chen0Yuhan Cheng1Liushi Meng2Jian Chen3Science Center for Future Foods, Jiangnan University, Wuxi 214122, ChinaScience Center for Future Foods, Jiangnan University, Wuxi 214122, ChinaJiaxing Synbiolab Technology Co., Ltd., Jiaxing 314000, ChinaScience Center for Future Foods, Jiangnan University, Wuxi 214122, ChinaTrehalose is crucial for the stress resistance of <i>Saccharomyces cerevisiae</i>, primarily through its stabilization of proteins and membranes. The Agt1 transporter, a member of the Major Facilitator Superfamily, mediates trehalose uptake, a key process for maintaining cellular integrity under stress. Despite its importance, the molecular mechanisms of Agt1-mediated trehalose transport remain underexplored. In this study, we expressed and purified the trehalase enzyme TreA from <i>E. coli</i> to develop reliable trehalose assays. We screened 257 wild <i>S. cerevisiae</i> isolates, identifying strains with enhanced trehalose transport capacities. Comparative analyses, including structural modeling and molecular docking, revealed that specific Agt1 variants exhibited significantly higher transport efficiency, influenced by key residues in the transporter. Molecular dynamics simulations and steered molecular dynamics provided further insights, particularly into the role of the Agt1 channel head region in substrate recognition and binding. Site-directed mutagenesis validated these findings, showing that mutations at critical residues, such as 156Q, 164L, 256Q, 395E, 396R, and 507Y significantly reduced transport activity, while 137Q, 230T, and 514 N increased efficiency under certain conditions.https://www.mdpi.com/2309-608X/10/11/781trehaloseAgt1<i>Saccharomyces cerevisiae</i>molecular dynamicssite-directed mutagenesisstress resistance |
| spellingShingle | Anqi Chen Yuhan Cheng Liushi Meng Jian Chen Key Amino Acid Residues of the Agt1 Transporter for Trehalose Transport by <i>Saccharomyces cerevisiae</i> Journal of Fungi trehalose Agt1 <i>Saccharomyces cerevisiae</i> molecular dynamics site-directed mutagenesis stress resistance |
| title | Key Amino Acid Residues of the Agt1 Transporter for Trehalose Transport by <i>Saccharomyces cerevisiae</i> |
| title_full | Key Amino Acid Residues of the Agt1 Transporter for Trehalose Transport by <i>Saccharomyces cerevisiae</i> |
| title_fullStr | Key Amino Acid Residues of the Agt1 Transporter for Trehalose Transport by <i>Saccharomyces cerevisiae</i> |
| title_full_unstemmed | Key Amino Acid Residues of the Agt1 Transporter for Trehalose Transport by <i>Saccharomyces cerevisiae</i> |
| title_short | Key Amino Acid Residues of the Agt1 Transporter for Trehalose Transport by <i>Saccharomyces cerevisiae</i> |
| title_sort | key amino acid residues of the agt1 transporter for trehalose transport by i saccharomyces cerevisiae i |
| topic | trehalose Agt1 <i>Saccharomyces cerevisiae</i> molecular dynamics site-directed mutagenesis stress resistance |
| url | https://www.mdpi.com/2309-608X/10/11/781 |
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