The Centre of Molecular Structure (CMS) consists of three core facilities: i) Biophysical Measurements; ii) Crystallization and X-ray Diffraction; iii) Structural Mass spectrometry. The CMS thus provides complementary tools for the studies of the biophysical properties, three-dimensional structure and structure:function relationships of biological macromolecules.
The Biophysical Techniques facility offers circular dichroism spectroscopy (Chirascan Plus CD spectrometer), spectrophotometry (Specord 50 Plus UV/Vis spectrophotometer), differential scanning fluorescence (Prometheus NT.48), dynamic light scattering (Zetasizer Nano ZS90) and differential scanning calorimetry (Microcal VP-DSC). These devices are used for the determination of size, structure and stability of biomolecules, and for the study of conformational changes and thermodynamics of temperature transitions. Isothermal titration calorimetry (Microcal iTC200), microscale thermophoresis (Monolith NT.115 and NT.LabelFree) and surface plasmon resonance (ProteOn XPR36) allow to characterize protein-protein interactions and protein-ligand interactions (small molecule, DNA, RNA, peptides, sugars, lipids…) that can be measured under close to native conditions.
The Crystallisation and Diffraction facility includes an ArtRobbins Gryphon dropsetter, which allows to set-up nanodrop crystallisation plates, and a Formulatrix RI1000 hotel for the storage of crystallization plates and for the automated, remote-access monitoring of crystallization droplets using visible, polarized and UV light. A glovebox, a cold room and a warm room (all equipped with stereomicroscopes) allow crystallisation and crystal manipulation in different environments (including in oxygen-free conditions). The Bruker D8 Venture diffractometer equipped with a liquid metal jet X-ray source (plus its ISX stage for in situ diffraction) is the core equipment for diffraction data acquisition.
The Structural Mass Spectrometry facility offers access to an ultra-high resolution 15T-Solarix XR FT-ICR (Bruker Daltonics), and to a MALDI-TOF Autoflex Speed (Bruker Daltonics) mass spectrometers. This instrumentation allows determination of precise mass of biological macromolecules, characterization of posttranslational modifications, peptide mass fingerprinting, detection of small molecule/metabolites and monitoring of protein structural changes/protein-protein interactions under physiological conditions by hydrogen-deuterium exchange and chemical cross-linking. An Agilent Technologies 1200 HPLC system coupled to the 15T-SolariX XR mass spectrometer is used for the separation of complex peptide mixtures, proteins and metabolites.
Together with the core facilities of CEITEC, the CMS is part of Czech Infrastructure for Integrative Structural Biology (CIISB), national affiliated centre of Instruct-ERIC (European Integrated Structural Biology Infrastructure).
This work was supported by the ERDF (BIOCEV: CZ.1.05/1.1.00/02.0109), MEYS CR (CIISB: LM2015043).