Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser
- Paul Scherrer Inst., Villigen (Switzerland). Lab. for Biomolecular Research
Rhodopsin is a membrane protein from the G protein-coupled receptor family. Together with its ligand retinal, it forms the visual pigment responsible for night vision. In order to perform ultrafast dynamics studies, a time-resolved serial femtosecond crystallography method is required owing to the nonreversible activation of rhodopsin. In such an approach, microcrystals in suspension are delivered into the X-ray pulses of an X-ray free-electron laser (XFEL) after a precise photoactivation delay. Here in this study, a millilitre batch production of high-density microcrystals was developed by four methodical conversion steps starting from known vapour-diffusion crystallization protocols: (i) screening the low-salt crystallization conditions preferred for serial crystallography by vapour diffusion, (ii) optimization of batch crystallization, (iii) testing the crystal size and quality using second-harmonic generation (SHG) imaging and X-ray powder diffraction and (iv) production of millilitres of rhodopsin crystal suspension in batches for serial crystallography tests; these crystals diffracted at an XFEL at the Linac Coherent Light Source using a liquid-jet setup.
- Research Organization:
- Paul Scherrer Inst., Villigen (Switzerland). Lab. for Biomolecular Research
- Sponsoring Organization:
- USDOE; National Institutes of Health (NIH); National Science Foundation (NSF)
- OSTI ID:
- 1345038
- Journal Information:
- Acta Crystallographica. Section F, Structural Biology Communications, Vol. 71, Issue 7; ISSN 2053-230X
- Publisher:
- International Union of CrystallographyCopyright Statement
- Country of Publication:
- United States
- Language:
- English
Web of Science
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