Reaction intermediates during DNA synthesis have been studied in detail using time-resolved
X-ray crystallography for translesion and repair DNA polymerases. Contrary to the originally
proposed two-metal-ion mechanism, a third metal ion was identified between the finger
domain and the α- and β-phosphates of the incoming nucleotide. This third metal ion was
suggested to either participate in...
Glucuronyl esterases (GEs) from the carbohydrate esterase family 15 (CE15) are involved in degrading lignocellulosic biomass, by catalyzing the hydrolysis of an esterbond connecting lignin and hemicellulose in the plant cell wall (1). In order to utilize biomass in biorefineries, efficient methods are needed to separate cellulose, hemicellulose and lignin. Studying GEs to better understand...
Serial crystallography has opened new possibilities for determining the structures of biological macromolecules using microcrystals. At the European XFEL, the Sample Environment and Characterization (SEC) group, operating the XBI biology laboratory, provides essential support for users at every stage of the serial crystallography workflow, from initial sample preparation to advanced...
To obtain the highest quality electron density maps, from a time-resolved serial crystallography experiment, it is crucial to accurately group the observed intensities according to the structure of the protein that generated those intensities. In an ideal world, each unique protein crystal structure would give rise to a set of well defined intensities, which would enable the intensities to be...
A highly flexible protein with an active site that changes its volume to fit a wide variety of ligands. A lid made of loops changes conformation based on ligand-size. Inhibition of this enzyme stops metabolism of drugs. This is an automatic disqualification of a drug candidate. Room temperature SSX shows better definition of some flexible loops, even at worse resolution. SSX data collection at...
The ALBA Synchrotron, the 3 GeV light source in Southern Europe, is preparing for its upgrade to a 4th generation storage ring (ALBA II) together with the construction of 3 beamlines and major upgrades on the existing ones, early next decade. Within this framework, serial and time-resolved macromolecular crystallography (SSX and TR-MX) are identified as strategic growth areas.
BL13-XALOC, in...
The X-ray Laue diffraction captures the crystal diffraction from white beam and can contain significant amount of structural information in comparison to monochromatic diffraction. In time-resolved study, the serial femtosecond crystallography (SFX) and serial synchrotron crystallography (SSX) are the mainstream approaches with the caveat of generating large data volume. Laue diffraction has...
Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that degrade polysaccharides oxidatively, with applications in second-generation bioethanol production and as virulence factors in certain pathogens [1] [2]. They have been reclassified within the CAZy database into auxiliary activity families AA9–AA11 and AA13–AA17 [3], and their active-site histidine brace is highly...
Metalloenzymes containing heme centres catalyse a wide range of reactions critical to life. Understanding the structure and electronic states of the heme centre across multiple functionally relevant states is essential to understand mechanism. I will describe work using time resolved serial crystallography as well as the use of X-ray dose for the manipulation of heme iron oxidation states in...
Serial crystallography (SX) has been a revolutionary technique in structural biology for more than a decade, providing insights into the structures and dynamics of biomolecules at room temperature. Using intense ultra-short X-ray pulses, SX made the collection of diffraction data from micron-sized crystals possible, avoiding radiation damage and allowing for the capture of transient states and...
Spectroscopic techniques provide a powerful means of obtaining detailed information on the structural and dynamic properties of proteins in solution and in crystallo (proteins and enzymes are active in the crystalline state). Structural data obtained by X-ray crystallography is strengthened by insights from complementary approaches like UV/vis, fluorescence and (resonance) Raman measurements.
The rise of 4th generation sources, including the MAX IV Laboratory 3 GeV ring, has enabled new possibilities to study dynamics using crystallography. The MicroMAX beamline is a new beamline focussed on providing optimal X-ray characteristics for serial (SSX) and time-resolved (TR-SSX) crystallography at MAX IV [1]. The beamline emphasizes a flexible sample environment for standard and bespoke...
Polyurethane (PU) plastics are extermyl durable and hard to recycle by convential methods due to
crosslinked and branched molecular structure. Recent discoveries of enzyme that arget carbamate
(urethane) bond in the PU provide alternative way of PU recycling by means of biocatalytic degradation.
EnZync center has been established to discover, characterize and engineer novel enzymes for...
