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Instruments / Technical Area	Publication Details	Published	Actions
I04-1-Macromolecular Crystallography (fixed wavelength) I04-Macromolecular Crystallography I24-Microfocus Macromolecular Crystallography	Identification and characterization of diverse OTU deubiquitinases in bacteria Alexander F. Schubert , Justine V Nguyen , Tyler G. Franklin , Paul P. Geurink , Cameron G. Roberts , Daniel J. Sanderson , Lauren N. Miller , Huib Ovaa , Kay Hofmann , Jonathan N. Pruneda , David Komander The Embo Journal 81 DOI: 10.15252/embj.2020105127 Diamond Proposal Number(s): [8547, 11235] Open Access Show Abstract ▼ Abstract: Manipulation of host ubiquitin signaling is becoming an increasingly apparent evolutionary strategy among bacterial and viral pathogens. By removing host ubiquitin signals, for example, invading pathogens can inactivate immune response pathways and evade detection. The ovarian tumor (OTU) family of deubiquitinases regulates diverse ubiquitin signals in humans. Viral pathogens have also extensively co‐opted the OTU fold to subvert host signaling, but the extent to which bacteria utilize the OTU fold was unknown. We have predicted and validated a set of OTU deubiquitinases encoded by several classes of pathogenic bacteria. Biochemical assays highlight the ubiquitin and polyubiquitin linkage specificities of these bacterial deubiquitinases. By determining the ubiquitin‐bound structures of two examples, we demonstrate the novel strategies that have evolved to both thread an OTU fold and recognize a ubiquitin substrate. With these new examples, we perform the first cross‐kingdom structural analysis of the OTU fold that highlights commonalities among distantly related OTU deubiquitinases.	Jun 2020
I18-Microfocus Spectroscopy	Transition metal distribution in the brain and spinal cord of a dysmyelinated rodent Alia Al-Ebraheem , Lauren Miller , Kalotina Geraki , Kimberly L. Desmond , Greg J. Stanisz , Nicholas A. Bock , Michael Farquharson ICXOM23 DOI: 10.1063/1.4961139 Diamond Proposal Number(s): [9704] Open Access Show Abstract ▼ Abstract: Transition metal concentrations in the central nervous system (CNS) are altered in neurodegenerative diseases such as Alzheimer’s, Parkinson’s and multiple sclerosis. A common symptom of these diseases is demyelination, which is the degradation of the myelin sheath that encapsulates the neurons in vertebrates. Transition metal concentrations were measured in Long Evans Shaker (LES) rodent model and compared to healthy age-matched controls to investigate the relationship between transition metals and myelination. Micro probe Synchrotron Radiation X-ray Fluorescence (µSRXRF) was used to measure concentrations of manganese (Mn), iron (Fe), copper (Cu), and zinc (Zn) in regions of grey matter and white matter in Shaker rodents and their age-matched Long Evans (LE) controls in the cerebellum and spinal cord. In the cerebellum, the concentrations of all elements were significantly increased in the white matter of the Shaker model, and decreased in the gray matter of the Shaker model in comparison to their age and region matched controls. In the spinal cord samples, concentrations of all metals were higher in white matter and grey matter of Shaker rat spinal cord compared to those in the control rat spinal cord. This study demonstrated that the sensitivity of µSRXRF is sufficient to discriminate between the elemental distributions of gray and white matter of the brain sections and spinal cords in the two groups. The observed significant increase of Mn, Fe, Zn and Cu in the white matter of the Shaker animals in the cerebellum and spinal cord compared to controls could be the result of astrocytic glial cells replacing the myelin in the CNS. Unlike other imaging techniques, the fine resolution of µSRXRF enables specific regions of gray matter structures namely, the molecular layer and the granule layer to be identified in the rat CNS, and their transition metal concentrations to be quantified. This work will further establish µSRXRF as a powerful analytic technique for compositional studies in brain sections from models of brain disease.	Sep 2016

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