VP28 interacts with PmRab7 irrespective of its nucleotide state
Abstract In shrimp aquaculture, white spot syndrome virus (WSSV) infections severely impact production. Previous research highlighted the crucial role of the Penaeus monodon Rab7 (PmRab7) protein in WSSV entry, specifically its interaction with the viral envelope protein VP28. PmRab7 exists in two c...
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Nature Portfolio
2024-11-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-79310-5 |
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| author | Patcha Sudsat Jiraporn Srisala Danaya Pakotiprapha Satita Tapaneeyakorn Kallaya Sritunyalucksana Siripong Thitamadee Sitthivut Charoensutthivarakul Ornchuma Itsathitphaisarn |
| author_facet | Patcha Sudsat Jiraporn Srisala Danaya Pakotiprapha Satita Tapaneeyakorn Kallaya Sritunyalucksana Siripong Thitamadee Sitthivut Charoensutthivarakul Ornchuma Itsathitphaisarn |
| author_sort | Patcha Sudsat |
| collection | DOAJ |
| description | Abstract In shrimp aquaculture, white spot syndrome virus (WSSV) infections severely impact production. Previous research highlighted the crucial role of the Penaeus monodon Rab7 (PmRab7) protein in WSSV entry, specifically its interaction with the viral envelope protein VP28. PmRab7 exists in two conformations: GDP-bound (inactive) and GTP-bound (active). This study, using ELISA and isothermal titration calorimetry (ITC), reveals that the PmRab7-VP28 interaction occurs irrespective of the nucleotide binding state of PmRab7. Comparing the binding affinity between VP28 and different PmRab7 conformations, including wild-type (WT, 22.5 nM), a fast nucleotide exchange (L129F, 128 nM), a GDP-bound form (T22N, 334 nM), and a favorably GTP-bound form (Q67L, 1990 nM), PmRab7-WT exhibits the strongest binding affinity, especially at a lower temperature (25 °C). The binding of PmRab7-WT and VP28 in the presence of excess nucleotide (WT with excess GDP, 924 nM, and WT with excess GTP, 826 nM) shows a 2-fold higher binding affinity than in the absence (WT, 1920 nM) indicating that the addition of excess nucleotide for PmRab7-WT enhanced the affinity for VP28. Together, these findings support the potential of PmRab7-WT as a promising therapeutic candidate for WSSV control in shrimp. Furthermore, from an industrial point of view, the ITC platform developed to study the VP28-PmRab7 interactions provides a high-throughput method for screening additives for shrimp feed that can inhibit this interaction. |
| format | Article |
| id | doaj-art-c44c99ac1c984cec8288583dda411c40 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-c44c99ac1c984cec8288583dda411c402024-11-17T12:27:40ZengNature PortfolioScientific Reports2045-23222024-11-0114111610.1038/s41598-024-79310-5VP28 interacts with PmRab7 irrespective of its nucleotide statePatcha Sudsat0Jiraporn Srisala1Danaya Pakotiprapha2Satita Tapaneeyakorn3Kallaya Sritunyalucksana4Siripong Thitamadee5Sitthivut Charoensutthivarakul6Ornchuma Itsathitphaisarn7Department of Biochemistry, Faculty of Science, Mahidol UniversityNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA)Department of Biochemistry, Faculty of Science, Mahidol UniversityNational Nanotechnology Center (NANOTEC), National Science and Technology Development Agency (NSTDA)National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA)Center of Excellence in Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol UniversitySchool of Bioinnovation and Bio-based Product Intelligence, Faculty of Science, Mahidol UniversityDepartment of Biochemistry, Faculty of Science, Mahidol UniversityAbstract In shrimp aquaculture, white spot syndrome virus (WSSV) infections severely impact production. Previous research highlighted the crucial role of the Penaeus monodon Rab7 (PmRab7) protein in WSSV entry, specifically its interaction with the viral envelope protein VP28. PmRab7 exists in two conformations: GDP-bound (inactive) and GTP-bound (active). This study, using ELISA and isothermal titration calorimetry (ITC), reveals that the PmRab7-VP28 interaction occurs irrespective of the nucleotide binding state of PmRab7. Comparing the binding affinity between VP28 and different PmRab7 conformations, including wild-type (WT, 22.5 nM), a fast nucleotide exchange (L129F, 128 nM), a GDP-bound form (T22N, 334 nM), and a favorably GTP-bound form (Q67L, 1990 nM), PmRab7-WT exhibits the strongest binding affinity, especially at a lower temperature (25 °C). The binding of PmRab7-WT and VP28 in the presence of excess nucleotide (WT with excess GDP, 924 nM, and WT with excess GTP, 826 nM) shows a 2-fold higher binding affinity than in the absence (WT, 1920 nM) indicating that the addition of excess nucleotide for PmRab7-WT enhanced the affinity for VP28. Together, these findings support the potential of PmRab7-WT as a promising therapeutic candidate for WSSV control in shrimp. Furthermore, from an industrial point of view, the ITC platform developed to study the VP28-PmRab7 interactions provides a high-throughput method for screening additives for shrimp feed that can inhibit this interaction.https://doi.org/10.1038/s41598-024-79310-5 |
| spellingShingle | Patcha Sudsat Jiraporn Srisala Danaya Pakotiprapha Satita Tapaneeyakorn Kallaya Sritunyalucksana Siripong Thitamadee Sitthivut Charoensutthivarakul Ornchuma Itsathitphaisarn VP28 interacts with PmRab7 irrespective of its nucleotide state Scientific Reports |
| title | VP28 interacts with PmRab7 irrespective of its nucleotide state |
| title_full | VP28 interacts with PmRab7 irrespective of its nucleotide state |
| title_fullStr | VP28 interacts with PmRab7 irrespective of its nucleotide state |
| title_full_unstemmed | VP28 interacts with PmRab7 irrespective of its nucleotide state |
| title_short | VP28 interacts with PmRab7 irrespective of its nucleotide state |
| title_sort | vp28 interacts with pmrab7 irrespective of its nucleotide state |
| url | https://doi.org/10.1038/s41598-024-79310-5 |
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