Rapid and efficient ambient temperature X-ray crystal structure determination at Turkish Light Source

Abstract

High-resolution biomacromolecular structure determination is essential to better understand protein function and dynamics. Serial crystallography is an emerging structural biology technique which has fundamental limitations due to either sample volume requirements or immediate access to the competitive X-ray beamtime. Obtaining a high volume of well-diffracting, sufficient-size crystals while mitigating radiation damage remains a critical bottleneck of serial crystallography. As an alternative, we introduce the plate-reader module adapted for using a 72-well Terasaki plate for biomacromolecule structure determination at a convenience of a home X-ray source. We also present the first ambient temperature lysozyme structure determined at the Turkish light source (Turkish DeLight). The complete dataset was collected in 18.5 min with resolution extending to 2.39 Å and 100% completeness. Combined with our previous cryogenic structure (PDB ID: 7Y6A), the ambient temperature structure provides invaluable information about the structural dynamics of the lysozyme. Turkish DeLight provides robust and rapid ambient temperature biomacromolecular structure determination with limited radiation damage.

X-ray Crystallographic and Hydrogen Deuterium Exchange Studies Confirm Alternate Kinetic Models for Homolog Insulin Monomers

“…Data collection was performed using Rigaku’s XtaLAB Synergy Flow X-ray diffraction (XRD) system, controlled by CrysAlisPro software (Rigaku Oxford Diffraction, 2022), following the protocol outlined in Gul et al (2023). The airflow temperature of Oxford Cryosystems’s Cryostream 800 Plus was set to 300 K (26.85 °C) and maintained consistently at ambient temperature during data collection.…”

Structural Analysis of Wild-type and Val120Thr MutantCandida boidiniiFormate Dehydrogenase by X-ray Crystallography

“…The harvested crystals were flash-frozen by plunging them in liquid nitrogen and placed in a cryo-cooled robotic sample puck (Cat#M-CP-111-021, MiTeGen, USA). The puck was mounted to a puck transfer and mounting tool and placed into the sample dewar which is auto refilled with liquid nitrogen at 100 °K, of the Turkish DeLight XRD device (Gul et al, 2023).…”

“…Data collection was performed for 15 hours 43 minutes 19 seconds. Data collection at ambient temperature was performed as described byGul et al (2023). Data reduction was performed using the CrysAlis Pro version 1.171.42.35a.…”

The structural studies of Escherichia coli CyaY at ambient and cryogenic temperatures

“…The collected data was then merged using pro t merge process with CrysAlisPro 1.171.42.59a software (Rigaku OxfordDiffraction,2022) to produce an integrated re ection dataset (*.mtz) le for further analysis, as described in Gul et.al. 2022 [42].…”

“…In addition, they have failed to show structural and functional heterogeneity of the complexes involved in cluster biogenesis. To address these limitations, we determined the X-ray crystal structure of CyaY from E. coli at ambient temperature (hereafter referred to as CyaY AT1 ) using our Turkish DeLight home X-ray source [41,42]. To perform a rigorous comparison and assess the effect of temperature, we also obtained the cryogenic crystal structure of CyaY (hereafter referred to as CyaY CT1 ) under the same experimental conditions.…”