Ice diffraction is a much bigger and more widespread problem in protein crystallography than commonly recognized [1]. 'Ice rings' are notoriously difficult to account for in integration [2]. The resulting experimental structure factor errors can affect the modelling of macromolecular structures and may even prevent their solution. Furthermore, ice rings are hard to recognize from diffraction...
EMBL-Hamburg operates two beamlines for macromolecular crystallography, P13 and P14, at PETRA III (DESY, Hamburg).
P13 delivers high photon fluxes at energies down to 4 keV. Combining X-rays in the 4-6 keV energy range with beam sizes down to 15 µm diameter while maintaining high photon flux and using standard mounting systems (SPINE pins) and robotics allows to solve the crystallographic...
The Helmholtz-Zentrum Berlin (HZB) operates three beamlines for macromolecular crystallography (MX) at the electron storage ring BESSY II [1,2]. BL14.1 and BL14.2 are tunable in the photon energy range from 5 to 16 keV, while BL14.3 is a fixed-energy side station (13.8 keV). They feature state-of-the-art experimental stations and ancillary facilities, serving more than 100 research groups...
A major success story of the European Synchrotron Radiation Facility (ESRF, the European Synchrotron) has been its facilities for Structural Biology. Here, end-stations for experiments in Macromolecular Crystallography, BioSAXS and in crystallo spectroscopy have recently been supplemented by the installation of a high-end cryo-electron microscope. The very high level of productivity of these...
Fragment-based lead discovery has become increasingly popular to identify first leads for drug development. Advantage of using fragments is their low molecular weight, which leaves sufficient space for subsequent chemical optimization before the pharmaceutically reasonable limit of 500 g/mol is reached. However, the inherent low-binding affinity of fragments poses major challenge to current...
Native ion mobility mass spectrometry (MS) is a perfect tool to study protein complexes in a mass and conformation specific manner. Despite remarkable sensitivity and selectivity the structural resolution is limited in native MS. On the other hand, it allows monitoring structural transitions, which proteins and protein complexes undergo, e.g. during the viral lifecycle. However, such transient...
Although X-ray crystallography allowed the determination of the three-dimensional structure of fundamental biomolecules such as proteins and nucleic acids, neutron crystallography provides a powerful complement because hydrogen atoms can be visualized directly. Due to the lower flux of neutrons compared to that of X-rays, bigger crystals (~0.5 mm3) are required. So, the improvement of the...
Modern X-ray sources such as synchrotrons and free-electron lasers provide the possibility of ultra-fast data recording within micro-seconds. Small angle X-ray scattering (SAXS) on biological macromolecules is one tool for exploring the kinetic and dynamic of structural alteration of proteins during the working cycle. Advanced sample environments based on microfluidic devices allows handling...
P11 at PETRA III in Hamburg is dedicated to structural investigations of biological samples at different length scales. The beamline provides two state-of-the-art endstations: a crystallography experiment which is in user operation since 2013 [1] and an X-ray microscope which is utilizing tender X-rays between 2.4 and 10 keV and is currently under construction [2]. In future, a third...
Dynamically resolved, or better known time-resolved, structural studies of proteins rely on specific sample environments at both synchrotrons and XFELs due to two main requirements: the continuous delivery of fresh sample to the X-ray beam and the need to trigger the protein activity. The two most widely used methods for protein triggering are the diffusion of actuators by rapid mixing and...
