Cryogenic X-ray crystallographic studies of biomacromolecules at Turkish Light Source "<i>Turkish DeLight</i>"

Abstract

X-ray crystallography is a robust and powerful structural biology technique that provides high-resolution atomic structures of biomacromolecules. Scientists use this technique to unravel mechanistic and structural details of biological macromolecules (e.g., proteins, nucleic acids, protein complexes, protein-nucleic acid complexes, or large biological compartments). Since its inception, single-crystal cryocrystallography has never been performed in Türkiye due to the lack of a single-crystal X-ray diffractometer. The X-ray diffraction facility recently established at the University of Health Sciences, İstanbul, Türkiye will enable Turkish and international researchers to easily perform high-resolution structural analysis of biomacromolecules from single crystals. Here, we describe the technical and practical outlook of a state-of-the-art home-source X-ray, using lysozyme as a model protein. The methods and practice described in this article can be applied to any biological sample for structural studies. Therefore, this article will be a valuable practical guide from sample preparation to data analysis.

Structural Characterization of TRAF6 N-terminal for Therapeutic Uses

“…The TRAF6 crystals were harvested using MiTeGen MicroLoops attached to a magnetic wand [32] while being monitored under microscope [33]. The obtained crystals were flash frozen by plunging in liquid nitrogen and placed in a cryo-cooled sample storage puck (Cat#M-CP-111-021, MiTeGen, USA).…”

Ambient temperature crystal structure ofEscherichia coliCyaY protein displays alternate conformation

“…2022 [43]. The diffraction data was then merged and processed with CrysAlisPro 1.171.42.59a software (Rigaku OxfordDiffraction,2022), as described in Atalay et al 2022 [44].…”

Comparative Study of High-resolution LysB29(Nε-myristoyl) des(B30) Insulin Structures Display Novel Dynamic Causal Interrelations in Monomeric-Dimeric Motions

“…The harvested crystals were immediately flash-frozen by plunging in the liquid nitrogen and placed in a previously cryo-cooled sample puck (Cat#M-CP-111-021, MiTeGen, USA). The filled puck within the liquid nitrogen was transferred to the dewar of the Turkish DeLight for diffraction data collection [17].…”

“…Diffraction data was collected by using Rigaku's XtaLAB Synergy Flow XRD equipped with CrysAlisPro software (Agilent, 2014) as described in Atalay et al 2022 [17].…”

“…In this work, we present the high resolution detemir crystal structure determined to 1.7 Å resolution at the cryogenic temperature. Diffraction data collected at the Turkish Light Source (AKA Turkish DeLight) [17]. Additionally, we employed computational Gaussian Network Model (GNM) analysis that we aimed at investigating the differences of monomer:dimer equilibrium dynamics between our high-resolution cryogenic structure with previously published detemir structure (PDB ID: 1XDA).…”