Small-wedge synchrotron and serial XFEL datasets for Cysteinyl leukotriene GPCRs

Egor Marin,Aleksandra Luginina,Alexey Mishin,Vadim Cherezov,Sergey Bukhdruker,Anastasiia Gusach,Valentin Gordeliy,Elizaveta Lyapina,Vitaly Polovinkin,Kirill Kovalev,Polina Khorn,Valentin Borshchevskiy,Andrey Rogachev

doi:10.1038/s41597-020-00729-2

Abstract

Structural studies of challenging targets such as G protein-coupled receptors (GPCRs) have accelerated during the last several years due to the development of new approaches, including small-wedge and serial crystallography. Here, we describe the deposition of seven datasets consisting of X-ray diffraction images acquired from lipidic cubic phase (LCP) grown microcrystals of two human GPCRs, Cysteinyl leukotriene receptors 1 and 2 (CysLT1R and CysLT2R), in complex with various antagonists. Five datasets were collected using small-wedge synchrotron crystallography (SWSX) at the European Synchrotron Radiation Facility with multiple crystals under cryo-conditions. Two datasets were collected using X-ray free electron laser (XFEL) serial femtosecond crystallography (SFX) at the Linac Coherent Light Source, with microcrystals delivered at room temperature into the beam within LCP matrix by a viscous media microextrusion injector. All seven datasets have been deposited in the open-access databases Zenodo and CXIDB. Here, we describe sample preparation and annotate crystallization conditions for each partial and full datasets. We also document full processing pipelines and provide wrapper scripts for SWSX and SFX data processing.A Correction to this paper has been published: https://doi.org/10.1038/s41597-020-00759-w

Highlights

Cysteinyl leukotrienes, produced from arachidonic acid via the 5-lipooxygenase pathway, are pro-inflammatory mediators that modulate vascular permeability and immune response; they are involved in multiple disorders including asthma, cardiovascular diseases and cancer[1]
Cysteinyl leukotrienes elicit their action through two G protein-coupled receptors (GPCRs), CysLT1R and CysLT2R, that share 38% sequence identity[1]
CysLT2R is found in cardiovascular and brain tissues, with demonstrated involvement in ischemia and acute brain injuries[3,4], the role of this receptor remains controversial and poorly understood. Both CysLT1R and CysLT2R have been implicated in progression of various cancers[5,6,7,8], while the mutated form of CysLT2R with L129Q substitution has been associated with uveal melanoma[9,10]

Summary

Background & Summary

Cysteinyl leukotrienes, produced from arachidonic acid via the 5-lipooxygenase pathway, are pro-inflammatory mediators that modulate vascular permeability and immune response; they are involved in multiple disorders including asthma, cardiovascular diseases and cancer[1]. Cysteinyl leukotrienes elicit their action through two G protein-coupled receptors (GPCRs), CysLT1R and CysLT2R, that share 38% sequence identity[1]. Over the last few years, small-wedge synchrotron crystallography (SWSX) and serial femtosecond crystallography (SFX) have developed into powerful techniques, enabling high-resolution structure determination of many difficult to crystallize targets[12,13]. We describe all data processing steps, provide supporting code and intermediate results, aiming for reproducibility of deposited data processing

Methods

Findings

Code availability