title: Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser.
abstract: 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, 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.
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, OFLC/103, 5232 Villigen-PSI, Switzerland.
authors.forename: Wenting
authors.initials: W
authors.lastname: Wu
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, OFLC/103, 5232 Villigen-PSI, Switzerland.
authors.forename: Przemyslaw
authors.initials: P
authors.lastname: Nogly
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, OFLC/103, 5232 Villigen-PSI, Switzerland.
authors.forename: Jan
authors.initials: J
authors.lastname: Rheinberger
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, OFLC/103, 5232 Villigen-PSI, Switzerland.
authors.forename: Leonhard M
authors.initials: LM
authors.lastname: Kick
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, OFLC/103, 5232 Villigen-PSI, Switzerland.
authors.forename: Cornelius
authors.initials: C
authors.lastname: Gati
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, OFLC/103, 5232 Villigen-PSI, Switzerland.
authors.forename: Garrett
authors.initials: G
authors.lastname: Nelson
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, OFLC/103, 5232 Villigen-PSI, Switzerland.
authors.forename: Xavier
authors.initials: X
authors.lastname: Deupi
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, OFLC/103, 5232 Villigen-PSI, Switzerland.
authors.forename: Jörg
authors.initials: J
authors.lastname: Standfuss
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, OFLC/103, 5232 Villigen-PSI, Switzerland.
authors.forename: Gebhard
authors.initials: G
authors.lastname: Schertler
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, OFLC/103, 5232 Villigen-PSI, Switzerland.
authors.forename: Valérie
authors.initials: V
authors.lastname: Panneels
mesh_terms: D000818:Animals; D002417:Cattle; D003460:Crystallization; D007834:Lasers / Q000706:statistics & numerical data*; D012243:Rhodopsin / Q000737:chemistry*; D014961:X-Ray Diffraction / Q000379:methods*
chemical_list: D012243:Rhodopsin
keywords: FEL; GPCR; batch crystallization; dynamics; serial crystallography