Structural information about the $trans-to-cis$ isomerization mechanism of the photoswitchable fluorescent protein rsEGFP2 revealed by multiscale infrared transient absorption
| Autor: | Lucas M. Uriarte, Raffaele Vitale, Stanisław Niziński, Kyprianos Hadjidemetriou, Ninon Zala, Andras Lukacs, Gregory M. Greetham, Igor V. Sazanovich, Martin Weik, Cyril Ruckebusch, Stephen R. Meech, Michel Sliwa |
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| Přispěvatelé: | Laboratoire Avancé de Spectroscopie pour les Intéractions la Réactivité et l'Environnement - UMR 8516 (LASIRE), Institut de Chimie du CNRS (INC)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Department of Biophysics, Medical School, Central Laser Facility (CLF), STFC Rutherford Appleton Laboratory (RAL), Science and Technology Facilities Council (STFC)-Science and Technology Facilities Council (STFC), University of East Anglia [Norwich] (UEA), ANR-15-CE32-0004,BioXFEL,Caractérisation d'états intermédiaires de protéines fluorescentes en utilisant des lasers à électrons libres X et les spectroscopies UV-visible et infrarouge ultra-rapides(2015) |
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
MESH: Protein Conformation
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] MESH: Spectrophotometry Infrared MESH: Isomerism General Materials Science MESH: Luminescent Proteins Physical and Theoretical Chemistry MESH: Benzylidene Compounds MESH: Imidazoles |
| Zdroj: | Journal of Physical Chemistry Letters Journal of Physical Chemistry Letters, 2022, 13 (5), pp.1194-1202. ⟨10.1021/acs.jpclett.1c02920⟩ |
| ISSN: | 1948-7185 |
| DOI: | 10.1021/acs.jpclett.1c02920⟩ |
| Popis: | International audience; RsEGFP2 is a reversibly photoswitchable fluorescent protein used in super-resolved optical microscopies, which can be toggled between a fluorescent On state and a nonfluorescent Off state. Previous time-resolved ultraviolet-visible spectroscopic studies have shown that the Off-to-On photoactivation extends over the femto- to millisecond time scale and involves two picosecond lifetime excited states and four ground state intermediates, reflecting a trans-to-cis excited state isomerization, a millisecond deprotonation, and protein structural reorganizations. Femto- to millisecond time-resolved multiple-probe infrared spectroscopy (TRMPS-IR) can reveal structural aspects of intermediate species. Here we apply TRMPS-IR to rsEGFP2 and implement a Savitzky-Golay derivative analysis to correct for baseline drift. The results reveal that a subpicosecond twisted excited state precursor controls the trans-to-cis isomerization and the chromophore reaches its final position in the protein pocket within 100 ps. A new step with a time constant of 42 ns is reported and assigned to structural relaxation of the protein that occurs prior to the deprotonation of the chromophore on the millisecond time scale. |
| Databáze: | OpenAIRE |
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