X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature
The photochromic fluorescent protein Skylan-NS (Nonlinear Structured illumination variant mEos3.1H62L) is a reversibly photoswitchable fluorescent protein which has an unilluminated/ground state with an anionic and cis chromophore conformation and high fluorescence quantum yield. Photo-conversion wi...
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MDPI AG
2017-09-01
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| Series: | International Journal of Molecular Sciences |
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| author | Christopher D. M. Hutchison Violeta Cordon-Preciado Rhodri M. L. Morgan Takanori Nakane Josie Ferreira Gabriel Dorlhiac Alvaro Sanchez-Gonzalez Allan S. Johnson Ann Fitzpatrick Clyde Fare Jon P. Marangos Chun Hong Yoon Mark S. Hunter Daniel P. DePonte Sébastien Boutet Shigeki Owada Rie Tanaka Kensuke Tono So Iwata Jasper J. van Thor |
| author_facet | Christopher D. M. Hutchison Violeta Cordon-Preciado Rhodri M. L. Morgan Takanori Nakane Josie Ferreira Gabriel Dorlhiac Alvaro Sanchez-Gonzalez Allan S. Johnson Ann Fitzpatrick Clyde Fare Jon P. Marangos Chun Hong Yoon Mark S. Hunter Daniel P. DePonte Sébastien Boutet Shigeki Owada Rie Tanaka Kensuke Tono So Iwata Jasper J. van Thor |
| author_sort | Christopher D. M. Hutchison |
| collection | DOAJ |
| description | The photochromic fluorescent protein Skylan-NS (Nonlinear Structured illumination variant mEos3.1H62L) is a reversibly photoswitchable fluorescent protein which has an unilluminated/ground state with an anionic and cis chromophore conformation and high fluorescence quantum yield. Photo-conversion with illumination at 515 nm generates a meta-stable intermediate with neutral trans-chromophore structure that has a 4 h lifetime. We present X-ray crystal structures of the cis (on) state at 1.9 Angstrom resolution and the trans (off) state at a limiting resolution of 1.55 Angstrom from serial femtosecond crystallography experiments conducted at SPring-8 Angstrom Compact Free Electron Laser (SACLA) at 7.0 keV and 10.5 keV, and at Linac Coherent Light Source (LCLS) at 9.5 keV. We present a comparison of the data reduction and structure determination statistics for the two facilities which differ in flux, beam characteristics and detector technologies. Furthermore, a comparison of droplet on demand, grease injection and Gas Dynamic Virtual Nozzle (GDVN) injection shows no significant differences in limiting resolution. The photoconversion of the on- to the off-state includes both internal and surface exposed protein structural changes, occurring in regions that lack crystal contacts in the orthorhombic crystal form. |
| first_indexed | 2024-04-13T12:43:19Z |
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| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-13T12:43:19Z |
| publishDate | 2017-09-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-00f82ad034f34aaf80ff9d77075098032022-12-22T02:46:27ZengMDPI AGInternational Journal of Molecular Sciences1422-00672017-09-01189191810.3390/ijms18091918ijms18091918X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room TemperatureChristopher D. M. Hutchison0Violeta Cordon-Preciado1Rhodri M. L. Morgan2Takanori Nakane3Josie Ferreira4Gabriel Dorlhiac5Alvaro Sanchez-Gonzalez6Allan S. Johnson7Ann Fitzpatrick8Clyde Fare9Jon P. Marangos10Chun Hong Yoon11Mark S. Hunter12Daniel P. DePonte13Sébastien Boutet14Shigeki Owada15Rie Tanaka16Kensuke Tono17So Iwata18Jasper J. van Thor19Molecular Biophysics, Imperial College London, South Kensington Campus, London SW7 2AZ, UKMolecular Biophysics, Imperial College London, South Kensington Campus, London SW7 2AZ, UKProtein Crystallography Facility, Centre for Structural Biology, Flowers Building, Department of Life Sciences, Imperial College London, London SW7 2AZ, UKDepartment of Biological Sciences, Graduate School of Science, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032, JapanMolecular Biophysics, Imperial College London, South Kensington Campus, London SW7 2AZ, UKMolecular Biophysics, Imperial College London, South Kensington Campus, London SW7 2AZ, UKQuantum Optics and Laser Science Group, Blackett Laboratory, Imperial College, London SW7 2AZ, UKQuantum Optics and Laser Science Group, Blackett Laboratory, Imperial College, London SW7 2AZ, UKDiamond Light Source Ltd., Diamond House, Harwell Science & Innovation Campus, Didcot OX11 0DE, UKMolecular Biophysics, Imperial College London, South Kensington Campus, London SW7 2AZ, UKQuantum Optics and Laser Science Group, Blackett Laboratory, Imperial College, London SW7 2AZ, UKLCLS, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USALCLS, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USALCLS, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USALCLS, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USARIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Hyogo 679-5148, JapanRIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Hyogo 679-5148, JapanRIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Hyogo 679-5148, JapanRIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Hyogo 679-5148, JapanMolecular Biophysics, Imperial College London, South Kensington Campus, London SW7 2AZ, UKThe photochromic fluorescent protein Skylan-NS (Nonlinear Structured illumination variant mEos3.1H62L) is a reversibly photoswitchable fluorescent protein which has an unilluminated/ground state with an anionic and cis chromophore conformation and high fluorescence quantum yield. Photo-conversion with illumination at 515 nm generates a meta-stable intermediate with neutral trans-chromophore structure that has a 4 h lifetime. We present X-ray crystal structures of the cis (on) state at 1.9 Angstrom resolution and the trans (off) state at a limiting resolution of 1.55 Angstrom from serial femtosecond crystallography experiments conducted at SPring-8 Angstrom Compact Free Electron Laser (SACLA) at 7.0 keV and 10.5 keV, and at Linac Coherent Light Source (LCLS) at 9.5 keV. We present a comparison of the data reduction and structure determination statistics for the two facilities which differ in flux, beam characteristics and detector technologies. Furthermore, a comparison of droplet on demand, grease injection and Gas Dynamic Virtual Nozzle (GDVN) injection shows no significant differences in limiting resolution. The photoconversion of the on- to the off-state includes both internal and surface exposed protein structural changes, occurring in regions that lack crystal contacts in the orthorhombic crystal form.https://www.mdpi.com/1422-0067/18/9/1918XFELSFXrsFPSkylan-NSSACLALCLS |
| spellingShingle | Christopher D. M. Hutchison Violeta Cordon-Preciado Rhodri M. L. Morgan Takanori Nakane Josie Ferreira Gabriel Dorlhiac Alvaro Sanchez-Gonzalez Allan S. Johnson Ann Fitzpatrick Clyde Fare Jon P. Marangos Chun Hong Yoon Mark S. Hunter Daniel P. DePonte Sébastien Boutet Shigeki Owada Rie Tanaka Kensuke Tono So Iwata Jasper J. van Thor X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature International Journal of Molecular Sciences XFEL SFX rsFP Skylan-NS SACLA LCLS |
| title | X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature |
| title_full | X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature |
| title_fullStr | X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature |
| title_full_unstemmed | X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature |
| title_short | X-ray Free Electron Laser Determination of Crystal Structures of Dark and Light States of a Reversibly Photoswitching Fluorescent Protein at Room Temperature |
| title_sort | x ray free electron laser determination of crystal structures of dark and light states of a reversibly photoswitching fluorescent protein at room temperature |
| topic | XFEL SFX rsFP Skylan-NS SACLA LCLS |
| url | https://www.mdpi.com/1422-0067/18/9/1918 |
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