5FVG

Structure of IrisFP at 100 K.

5FVG の概要

• エントリーDOI: 10.2210/pdb5fvg/pdb
• 関連するPDBエントリー: 5FVF 5FVI
• 分子名称: Green to red photoconvertible GFP-like protein EosFP, SULFATE ION (3 entities in total)
• 機能のキーワード: fluorescent protein, flurescent protein
• 由来する生物種: Lobophyllia hemprichii (Lobed brain coral)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 104245.11

構造登録者

Colletier, J.P.,Gallat, F.X.,Coquelle, N.,Weik, M. (登録日: 2016-02-07, 公開日: 2017-01-11, 最終更新日: 2024-01-10)

主引用文献

Colletier, J.,Sliwa, M.,Gallat, F.,Sugahara, M.,Guillon, V.,Schiro, G.,Coquelle, N.,Woodhouse, J.,Roux, L.,Gotthard, G.,Royant, A.,Uriarte, L.M.,Ruckebusch, C.,Joti, Y.,Byrdin, M.,Mizohata, E.,Nango, E.,Tanaka, T.,Tono, K.,Yabashi, M.,Adam, V.,Cammarata, M.,Schlichting, I.,Bourgeois, D.,Weik, M.
Serial Femtosecond Crystallography and Ultrafast Absorption Spectroscopy of the Photoswitchable Fluorescent Protein Irisfp.
J.Phys.Chem.Lett, 7:882-, 2016

PubMed Abstract: Reversibly photoswitchable fluorescent proteins find growing applications in cell biology, yet mechanistic details, in particular on the ultrafast photochemical time scale, remain unknown. We employed time-resolved pump-probe absorption spectroscopy on the reversibly photoswitchable fluorescent protein IrisFP in solution to study photoswitching from the nonfluorescent (off) to the fluorescent (on) state. Evidence is provided for the existence of several intermediate states on the pico- and microsecond time scales that are attributed to chromophore isomerization and proton transfer, respectively. Kinetic modeling favors a sequential mechanism with the existence of two excited state intermediates with lifetimes of 2 and 15 ps, the second of which controls the photoswitching quantum yield. In order to support that IrisFP is suited for time-resolved experiments aiming at a structural characterization of these ps intermediates, we used serial femtosecond crystallography at an X-ray free electron laser and solved the structure of IrisFP in its on state. Sample consumption was minimized by embedding crystals in mineral grease, in which they remain photoswitchable. Our spectroscopic and structural results pave the way for time-resolved serial femtosecond crystallography aiming at characterizing the structure of ultrafast intermediates in reversibly photoswitchable fluorescent proteins.

PubMed: 26866390
DOI: 10.1021/ACS.JPCLETT.5B02789

実験手法

X-RAY DIFFRACTION (1.9 Å)