Dataset Information

Lipidic cubic phase serial femtosecond crystallography structure of a photosynthetic reaction centre.

ABSTRACT: Serial crystallography is a rapidly growing method that can yield structural insights from microcrystals that were previously considered to be too small to be useful in conventional X-ray crystallography. Here, conditions for growing microcrystals of the photosynthetic reaction centre of Blastochloris viridis within a lipidic cubic phase (LCP) crystallization matrix that employ a seeding protocol utilizing detergent-grown crystals with a different crystal packing are described. LCP microcrystals diffracted to 2.25 Å resolution when exposed to XFEL radiation, which is an improvement of 0.15 Å over previous microcrystal forms. Ubiquinone was incorporated into the LCP crystallization media and the resulting electron density within the mobile Q_B pocket is comparable to that of other cofactors within the structure. As such, LCP microcrystallization conditions will facilitate time-resolved diffraction studies of electron-transfer reactions to the mobile quinone, potentially allowing the observation of structural changes associated with the two electron-transfer reactions leading to complete reduction of the ubiquinone ligand.

SUBMITTER: Bath P

PROVIDER: S-EPMC9159286 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Lipidic cubic phase serial femtosecond crystallography structure of a photosynthetic reaction centre.

Båth Petra P Banacore Analia A Börjesson Per P Bosman Robert R Wickstrand Cecilia C Safari Cecilia C Dods Robert R Ghosh Swagatha S Dahl Peter P Ortolani Giorgia G Björg Ulfarsdottir Tinna T Hammarin Greger G García Bonete María José MJ Vallejos Adams A Ostojić Lucija L Edlund Petra P Linse Johanna Barbara JB Andersson Rebecka R Nango Eriko E Owada Shigeki S Tanaka Rie R Tono Kensuke K Joti Yasumasa Y Nureki Osamu O Luo Fangjia F James Daniel D Nass Karol K Johnson Philip J M PJM Knopp Gregor G Ozerov Dmitry D Cirelli Claudio C Milne Christopher C Iwata So S Brändén Gisela G Neutze Richard R

Acta crystallographica. Section D, Structural biology 20220525 Pt 6

Serial crystallography is a rapidly growing method that can yield structural insights from microcrystals that were previously considered to be too small to be useful in conventional X-ray crystallography. Here, conditions for growing microcrystals of the photosynthetic reaction centre of Blastochloris viridis within a lipidic cubic phase (LCP) crystallization matrix that employ a seeding protocol utilizing detergent-grown crystals with a different crystal packing are described. LCP microcrystals ...[more]

PMID: 35647917

Similar Datasets

Project description:Membrane proteins (MPs) are essential components of cellular membranes and primary drug targets. Rational drug design relies on precise structural information, typically obtained by crystallography; however MPs are difficult to crystallize. Recent progress in MP structural determination has benefited greatly from the development of lipidic cubic phase (LCP) crystallization methods, which typically yield well-diffracting, but often small crystals that suffer from radiation damage during traditional crystallographic data collection at synchrotron sources. The development of new-generation X-ray free-electron laser (XFEL) sources that produce extremely bright femtosecond pulses has enabled room temperature data collection from microcrystals with no or negligible radiation damage. Our recent efforts in combining LCP technology with serial femtosecond crystallography (LCP-SFX) have resulted in high-resolution structures of several human G protein-coupled receptors, which represent a notoriously difficult target for structure determination. In the LCP-SFX technique, LCP is recruited as a matrix for both growth and delivery of MP microcrystals to the intersection of the injector stream with an XFEL beam for crystallographic data collection. It has been demonstrated that LCP-SFX can substantially improve the diffraction resolution when only sub-10 µm crystals are available, or when the use of smaller crystals at room temperature can overcome various problems associated with larger cryocooled crystals, such as accumulation of defects, high mosaicity and cryocooling artifacts. Future advancements in X-ray sources and detector technologies should make serial crystallography highly attractive and practicable for implementation not only at XFELs, but also at more accessible synchrotron beamlines. Here we present detailed visual protocols for the preparation, characterization and delivery of microcrystals in LCP for serial crystallography experiments. These protocols include methods for conducting crystallization experiments in syringes, detecting and characterizing the crystal samples, optimizing crystal density, loading microcrystal laden LCP into the injector device and delivering the sample to the beam for data collection.

Dataset Information

Lipidic cubic phase serial femtosecond crystallography structure of a photosynthetic reaction centre.

Publications

Lipidic cubic phase serial femtosecond crystallography structure of a photosynthetic reaction centre.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets