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Title: Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser

Journal Article · · Acta Crystallographica. Section F, Structural Biology Communications

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
Citation Metrics:
Cited by: 12 works
Citation information provided by
Web of Science

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Cited By (10)

Protein crystal screening and characterization for serial femtosecond nanocrystallography journal May 2016
Time-resolved structural studies with serial crystallography: A new light on retinal proteins journal June 2015
Perspective: Opportunities for ultrafast science at SwissFEL journal November 2017
On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies text January 2019
Time-resolved structural studies with serial crystallography: A new light on retinal proteins text January 2015
Perspective: Opportunities for ultrafast science at SwissFEL text January 2017
Strategies for sample delivery for femtosecond crystallography text January 2019
On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies text January 2019
Perspective: Opportunities for ultrafast science at SwissFEL text January 2017
Protein crystal screening and characterization for serial femtosecond nanocrystallography text January 2023

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