A metastable prerequisite for the growth of lumazine synthase crystals

Dense liquid phases, metastable with respect to a solid phase, form in solutions of proteins and small-molecule materials. They have been shown to serve as a prerequisite for the nucleation of crystals and other ordered solid phases. Here, using crystals of the protein lumazine synthase from Bacillus subtilis, which grow by the generation and spreading of layers, we demonstrate that within a range of supersaturations the only mechanism of generation of growth layers involves the association of submicrometer-size droplets of the dense liquid to the crystal surface. The dense liquid is metastable not only with respect to the crystals, but also with respect to the low-concentration solution: dynamic light scattering reveals that the droplets' lifetime is limited to several seconds, after which they decay into the low-concentration solution. The short lifetime does not allow growth to detectable dimensions so that liquid-liquid phase separation is not observed within a range of conditions broader than the one used for crystallization. If during their lifetime the droplets encounter a crystal surface, they lower their free energy not by decay, but by transformation into crystalline matter, ensuring perfect registry with the substrate. These observations illustrate two novel features of phase transformations in solutions: the existence of doubly metastable, short-lifetime dense phases and their crucial role for the growth of an ordered solid phase.     

Calcium sulfate hemihydrate is the inorganic mineral in statoliths of Scyphozoan medusae (Cnidaria)

Scyphomedusae use inorganic crystals (statoliths) for gravity sensing. The organs which contain the statoliths are called rhopalia. Rhopalia of five different species of the three different orders of the class Scyphozoa were studied with high-end solid-state chemical methods to elucidate the crystallographic nature of the biomineral: synchrotron powder diffraction, synchrotron single-crystal diffraction, synchrotron microtomography, scanning electron microscopy, and energy dispersive X-ray spectroscopy. Each rhopalium contains a large number of statoliths in an ordered way. The statoliths of all species consist of calcium sulfate hemihydrate, a water-deficient phase. This is remarkable for sea-living organisms consisting mostly of water. The phylogenetic relationships within the class Scyphozoa are discussed.     

A comparative study of fragment screening methods on the p38�� kinase: new methods, new insights

The stress-activated kinase p38α was used to evaluate a fragment-based drug discovery approach using the BioFocus fragment library. Compounds were screened by surface plasmon resonance (SPR) on a Biacore(?) T100 against p38α and two selectivity targets. A sub-set of our library was the focus of detailed follow-up analyses that included hit confirmation, affinity determination on 24 confirmed, selective hits and competition assays of these hits with respect to a known ATP binding site inhibitor. In addition, functional activity against p38α was assessed in a biochemical assay using a mobility shift platform (LC3000, Caliper LifeSciences). A selection of fragments was also evaluated using fluorescence lifetime (FLEXYTE(?)) and microscale thermophoresis (Nanotemper) technologies. A good correlation between the data for the different assays was found. Crystal structures were solved for four of the small molecules complexed to p38α. Interestingly, as determined both by X-ray analysis and SPR competition experiments, three of the complexes involved the fragment at the ATP binding site, while the fourth compound bound in a distal site that may offer potential as a novel drug target site. A first round of optimization around the remotely bound fragment has led to the identification of a series of triazole-containing compounds. This approach could form the basis for developing novel and active p38α inhibitors. More broadly, it illustrates the power of combining a range of biophysical and biochemical techniques to the discovery of fragments that facilitate the development of novel modulators of kinase and other drug targets.     

Orbifolds and Topological Defects

We study orbifolds of two-dimensional topological field theories using defects. If the TFT arises as the twist of a superconformal field theory, we recover results on the Neveu–Schwarz and Ramond sectors of the orbifold theory, as well as bulk-boundary correlators from a novel, universal perspective. This entails a structure somewhat weaker than ordinary TFT, which however still describes a sector of the underlying conformal theory. The case of B-twisted Landau–Ginzburg models is discussed in detail, where we compute charge vectors and superpotential terms for B-type branes. Our construction also works in the absence of supersymmetry and for generalised “orbifolds” that need not arise from symmetry groups. In general, this involves a natural appearance of Hochschild (co)homology in a 2-categorical setting, in which among other things we provide simple presentations of Serre functors and a further generalisation of the Cardy condition.