Molecular dynamics guided identification of a brighter variant of superfolder Green Fluorescent Protein with increased photobleaching resistance
Abstract Fluorescent proteins (FPs) are a crucial tool for cell imaging, but with developments in fluorescence microscopy and researcher requirements there is still a need to develop brighter versions that remain fluorescent for longer. Using short time-scale molecular dynamics-based modelling to pr...
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| Format: | Article |
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Nature Portfolio
2025-06-01
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| Series: | Communications Chemistry |
| Online Access: | https://doi.org/10.1038/s42004-025-01573-4 |
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| author | Rochelle D. Ahmed W. David Jamieson Danoo Vitsupakorn Athena Zitti Kai A. Pawson Oliver K. Castell Peter D. Watson D. Dafydd Jones |
| author_facet | Rochelle D. Ahmed W. David Jamieson Danoo Vitsupakorn Athena Zitti Kai A. Pawson Oliver K. Castell Peter D. Watson D. Dafydd Jones |
| author_sort | Rochelle D. Ahmed |
| collection | DOAJ |
| description | Abstract Fluorescent proteins (FPs) are a crucial tool for cell imaging, but with developments in fluorescence microscopy and researcher requirements there is still a need to develop brighter versions that remain fluorescent for longer. Using short time-scale molecular dynamics-based modelling to predict changes in local chromophore interaction networks and solvation, we constructed an Aequorea victoria GFP (avGFP) variant called YuzuFP that is 1.5 times brighter than the starting superfolding variant (sfGFP) with a near 3-fold increased resistance to photobleaching in situ. YuzuFP contained a single mutation that replaces the chromophore interacting residue H148 with a serine. Longer time scale molecular dynamics revealed the likely mechanism of action: S148 forms more persistent H-bond with the chromophore phenolate group and increases the residency time of an important water molecule. As demonstrated by live cell imaging, YuzuFP not only offers a timely upgrade as a useful green-yellow avGFP for cell imaging applications over longer time scales, but it also provides a basic scaffold for future avGFP engineering efforts. |
| format | Article |
| id | doaj-art-2162e4e948544249b420394ac3aab042 |
| institution | Kabale University |
| issn | 2399-3669 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Nature Portfolio |
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| series | Communications Chemistry |
| spelling | doaj-art-2162e4e948544249b420394ac3aab0422025-08-20T03:25:15ZengNature PortfolioCommunications Chemistry2399-36692025-06-018111110.1038/s42004-025-01573-4Molecular dynamics guided identification of a brighter variant of superfolder Green Fluorescent Protein with increased photobleaching resistanceRochelle D. Ahmed0W. David Jamieson1Danoo Vitsupakorn2Athena Zitti3Kai A. Pawson4Oliver K. Castell5Peter D. Watson6D. Dafydd Jones7Molecular Bioscience Division, School of Biosciences, Cardiff UniversitySchool of Pharmacy and Pharmaceutical Sciences, Cardiff UniversityMolecular Bioscience Division, School of Biosciences, Cardiff UniversityMolecular Bioscience Division, School of Biosciences, Cardiff UniversityMolecular Bioscience Division, School of Biosciences, Cardiff UniversitySchool of Pharmacy and Pharmaceutical Sciences, Cardiff UniversityMolecular Bioscience Division, School of Biosciences, Cardiff UniversityMolecular Bioscience Division, School of Biosciences, Cardiff UniversityAbstract Fluorescent proteins (FPs) are a crucial tool for cell imaging, but with developments in fluorescence microscopy and researcher requirements there is still a need to develop brighter versions that remain fluorescent for longer. Using short time-scale molecular dynamics-based modelling to predict changes in local chromophore interaction networks and solvation, we constructed an Aequorea victoria GFP (avGFP) variant called YuzuFP that is 1.5 times brighter than the starting superfolding variant (sfGFP) with a near 3-fold increased resistance to photobleaching in situ. YuzuFP contained a single mutation that replaces the chromophore interacting residue H148 with a serine. Longer time scale molecular dynamics revealed the likely mechanism of action: S148 forms more persistent H-bond with the chromophore phenolate group and increases the residency time of an important water molecule. As demonstrated by live cell imaging, YuzuFP not only offers a timely upgrade as a useful green-yellow avGFP for cell imaging applications over longer time scales, but it also provides a basic scaffold for future avGFP engineering efforts.https://doi.org/10.1038/s42004-025-01573-4 |
| spellingShingle | Rochelle D. Ahmed W. David Jamieson Danoo Vitsupakorn Athena Zitti Kai A. Pawson Oliver K. Castell Peter D. Watson D. Dafydd Jones Molecular dynamics guided identification of a brighter variant of superfolder Green Fluorescent Protein with increased photobleaching resistance Communications Chemistry |
| title | Molecular dynamics guided identification of a brighter variant of superfolder Green Fluorescent Protein with increased photobleaching resistance |
| title_full | Molecular dynamics guided identification of a brighter variant of superfolder Green Fluorescent Protein with increased photobleaching resistance |
| title_fullStr | Molecular dynamics guided identification of a brighter variant of superfolder Green Fluorescent Protein with increased photobleaching resistance |
| title_full_unstemmed | Molecular dynamics guided identification of a brighter variant of superfolder Green Fluorescent Protein with increased photobleaching resistance |
| title_short | Molecular dynamics guided identification of a brighter variant of superfolder Green Fluorescent Protein with increased photobleaching resistance |
| title_sort | molecular dynamics guided identification of a brighter variant of superfolder green fluorescent protein with increased photobleaching resistance |
| url | https://doi.org/10.1038/s42004-025-01573-4 |
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