Crystal Structure of the N-Acetylmannosamine Kinase Domain of GNE

UDP-GlcNAc 2-epimerase/ManNAc 6-kinase, GNE, is a bi-functional enzyme that plays a key role in sialic acid biosynthesis. Mutations of the GNE protein cause sialurea or autosomal recessive inclusion body myopathy/Nonaka myopathy. GNE is the only human protein that contains a kinase domain belonging to the ROK (repressor, ORF, kinase) family.We solved the structure of the GNE kinase domain in the ligand-free state. The protein exists predominantly as a dimer in solution, with small populations of monomer and higher-order oligomer in equilibrium with the dimer. Crystal packing analysis reveals the existence of a crystallographic hexamer, and that the kinase domain dimerizes through the C-lobe subdomain. Mapping of disease-related missense mutations onto the kinase domain structure revealed that the mutation sites could be classified into four different groups based on the location - dimer interface, interlobar helices, protein surface, or within other secondary structural elements.The crystal structure of the kinase domain of GNE provides a structural basis for understanding disease-causing mutations and a model of hexameric wild type full length enzyme.This article can also be viewed as an enhanced version in which the text of the article is integrated with interactive 3D representations and animated transitions. Please note that a web plugin is required to access this enhanced functionality. Instructions for the installation and use of the web plugin are available in Text S1

Formation of a Nematic Fluid at High Fields in Sr3Ru2O7

In principle, a complex assembly of strongly interacting electrons can self-organise into a wide variety of collective states, but relatively few such states have been identified in practice. We report that, in the close vicinity of a metamagnetic quantum critical point, high purity Sr3Ru2O7 possesses a large magnetoresistive anisotropy, consistent with the existence of an electronic nematic fluid. We discuss a striking phenomenological similarity between our observations and those made in high purity two-dimensional electron fluids in GaAs devices.Comment: 14 pages, 3 figures, 11 extra pages of supplementary informatio

Research Data Management in the Atlantic Provinces

Research Data Management (RDM) is becoming an increasingly important issue for post-secondary institutions. The Tri-Agencies (SSHRC, NSERC, CIHR) are expected to release a new policy that will stipulate that institutions have an institutional strategy for dealing with research data. Part of the responsibility will lie with researchers and the management of their research data. Three Atlantic universities are aiming to discover how researchers are storing, using and sharing their data through administering a survey using an instrument that has been used at many Canadian universities. Surveys will be administered in Winter 2019. Data from the surveys will be analyzed separately by each institution and may be combined to identify commonalities and possibilities for collaboration in the region. The results of these surveys will help librarians, IT specialists, data managers, and others interested in data management to understand current practices and attitudes. The results will also be used to contribute of the development of RDM services and resources across Atlantic post-secondary institutions with the aim of being prepared for the Tri-Agency policy once it comes into effect

Superconducting Magnetization above the Irreversibility Line in Tl2Ba2CuO6

Piezolever torque magnetometry has been used to measure the magnetization of superconducting Tl2Ba2CuO6. Three crystals with different levels of oxygen overdoping were investigated in magnetic fields up to 10 Tesla. In all cases, the magnetization above the irreversibility line was found to depart from the behaviour M ~ ln(Hc2/H) of a simple London-like vortex liquid. In particular, for a strongly overdoped (Tc = 15K) crystal, the remnant superconducting order above the irreversibility line is characterized by a linear diamagnetic response (M ~ H) that persists well above Tc and also up to the highest field employed.Comment: RevTeX, 11 pages, 7 encapsulated PostScript figures, submitted to Physical Review

Reprocessing with GANEX:Methodology for Ligand Radiation Tolerance Testing

Results demonstrating the methodology for testing the radiation tolerance of organic ligands are presented. A high activity sealed source was used to irradiate samples which were sequentially removed and analysed using a sensitive mass spectrometer. The degradation of a candidate ligand for a new reprocessing process “GANEX” was found to be around 50% after 567 kGy exposure to gamma from Cs-137.<br/

Crystal Structures of the Tetratricopeptide Repeat Domains of Kinesin Light Chains: Insight into Cargo Recognition Mechanisms

Kinesin-1 transports various cargos along the axon by interacting with the cargos through its light chain subunit. Kinesin light chains (KLC) utilize its tetratricopeptide repeat (TPR) domain to interact with over 10 different cargos. Despite a high sequence identity between their TPR domains (87%), KLC1 and KLC2 isoforms exhibit differential binding properties towards some cargos. We determined the structures of human KLC1 and KLC2 tetratricopeptide repeat (TPR) domains using X-ray crystallography and investigated the different mechanisms by which KLCs interact with their cargos. Using isothermal titration calorimetry, we attributed the specific interaction between KLC1 and JNK-interacting protein 1 (JIP1) cargo to residue N343 in the fourth TRP repeat. Structurally, the N343 residue is adjacent to other asparagines and lysines, creating a positively charged polar patch within the groove of the TPR domain. Whereas, KLC2 with the corresponding residue S328 did not interact with JIP1. Based on these finding, we propose that N343 of KLC1 can form “a carboxylate clamp” with its neighboring asparagine to interact with JIP1, similar to that of HSP70/HSP90 organizing protein-1's (HOP1) interaction with heat shock proteins. For the binding of cargos shared by KLC1 and KLC2, we propose a different site located within the groove but not involving N343. We further propose a third binding site on KLC1 which involves a stretch of polar residues along the inter-TPR loops that may form a network of hydrogen bonds to JIP3 and JIP4. Together, these results provide structural insights into possible mechanisms of interaction between KLC TPR domains and various cargo proteins

Whey Protein Ingestion Activates mTOR-dependent Signalling after Resistance Exercise in Young Men: A Double-Blinded Randomized Controlled Trial

The effect of resistance exercise with the ingestion of supplementary protein on the activation of the mTOR cascade, in human skeletal muscle has not been fully elucidated. In this study, the impact of a single bout of resistance exercise, immediately followed by a single dose of whey protein isolate (WPI) or placebo supplement, on the activation of mTOR signalling was analyzed. Young untrained men completed a maximal single-legged knee extension exercise bout and were randomized to ingest either WPI supplement (n = 7) or the placebo (n = 7). Muscle biopsies were taken from the vastus lateralis before, and 2, 4 and 24 h post-exercise. WPI or placebo ingestion consumed immediately post-exercise had no impact on the phosphorylation of Akt (Ser473). However, WPI significantly enhanced phosphorylation of mTOR (Ser2448), 4E-BP1 (Thr37/46) and p70S6K (Thr389) at 2 h post-exercise. This study demonstrates that a single dose of WPI, when consumed in modest quantities, taken immediately after resistance exercise elicits an acute and transient activation of translation initiation within the exercised skeletal muscle