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Instruments / Technical Area	Publication Details	Published
I04-Macromolecular Crystallography	Staphylococcal complement evasion protein Sbi stabilises C3d dimers by inducing an N-terminal helix swap Rhys W. Dunphy , Ayla A. Wahid , Catherine R. Back , Rebecca L. Martin , Andrew G. Watts , Charlotte A. Dodson , Susan J. Crennell , Jean M. H. Van Den Elsen Frontiers In Immunology 13 DOI: 10.3389/fimmu.2022.892234 Diamond Proposal Number(s): [23269] Open Access Show Abstract ▼ Abstract: Staphylococcus aureus is an opportunistic pathogen that is able to thwart an effective host immune response by producing a range of immune evasion molecules, including S. aureus binder of IgG (Sbi) which interacts directly with the central complement component C3, its fragments and associated regulators. Recently we reported the first structure of a disulfide-linked human C3d17C dimer and highlighted its potential role in modulating B-cell activation. Here we present an X-ray crystal structure of a disulfide-linked human C3d17C dimer, which undergoes a structurally stabilising N-terminal 3D domain swap when in complex with Sbi. These structural studies, in combination with circular dichroism and fluorescence spectroscopic analyses, reveal the mechanism underpinning this unique helix swap event and could explain the origins of a previously discovered N-terminally truncated C3dg dimer isolated from rat serum. Overall, our study unveils a novel staphylococcal complement evasion mechanism which enables the pathogen to harness the ability of dimeric C3d to modulate B-cell activation.	May 2022
I04-Macromolecular Crystallography	Insights into the structure-function relationships of dimeric C3d fragments Ayla A. Wahid , Rhys W. Dunphy , Alex Macpherson , Beth G. Gibson , Liudmila Kulik , Kevin Whale , Catherine Back , Thomas M. Hallam , Bayan Alkhawaja , Rebecca L. Martin , Ingrid Meschede , Maisem Laabei , Alastair D. G. Lawson , V. Michael Holers , Andrew G. Watts , Susan J. Crennell , Claire L. Harris , Kevin J. Marchbank , Jean M. H. Van Den Elsen Frontiers In Immunology 12 DOI: 10.3389/fimmu.2021.714055 Diamond Proposal Number(s): [17212] Open Access Show Abstract ▼ Abstract: Cleavage of C3 to C3a and C3b plays a central role in the generation of complement-mediated defences. Although the thioester-mediated surface deposition of C3b has been well-studied, fluid phase dimers of C3 fragments remain largely unexplored. Here we show C3 cleavage results in the spontaneous formation of C3b dimers and present the first X-ray crystal structure of a disulphide-linked human C3d dimer. Binding studies reveal these dimers are capable of crosslinking complement receptor 2 and preliminary cell-based analyses suggest they could modulate B cell activation to influence tolerogenic pathways. Altogether, insights into the physiologically-relevant functions of C3d(g) dimers gained from our findings will pave the way to enhancing our understanding surrounding the importance of complement in the fluid phase and could inform the design of novel therapies for immune system disorders in the future.	Aug 2021
B07-C-Versatile Soft X-ray beamline: Ambient Pressure XPS and NEXAFS	Ambient-pressure endstation of the Versatile Soft X-ray (VerSoX) beamline at Diamond Light Source Georg Held , Federica Venturini , David C. Grinter , Pilar Ferrer , Rosa Arrigo , Liam Deacon , Wilson Quevedo Garzon , Kanak Roy , Alex Large , Christopher Stephens , Andrew Watts , Paul Larkin , Matthew Hand , Hongchang Wang , Linda Pratt , James J. Mudd , Thomas Richardson , Suren Patel , Michael Hillman , Stewart Scott Journal Of Synchrotron Radiation 27 DOI: 10.1107/S1600577520009157 Open Access Show Abstract ▼ Abstract: The ambient-pressure endstation and branchline of the Versatile Soft X-ray (VerSoX) beamline B07 at Diamond Light Source serves a very diverse user community studying heterogeneous catalysts, pharmaceuticals and biomaterials under realistic conditions, liquids and ices, and novel electronic, photonic and battery materials. The instrument facilitates studies of the near-surface chemical composition, electronic and geometric structure of a variety of samples using X-ray photoelectron spectroscopy (XPS) and near-edge X-ray absorption fine-structure (NEXAFS) spectroscopy in the photon energy range from 170 eV to 2800 eV. The beamline provides a resolving power hν/Δ(hν) > 5000 at a photon flux > 1010 photons s−1 over most of its energy range. By operating the optical elements in a low-pressure oxygen atmosphere, carbon contamination can be almost completely eliminated, which makes the beamline particularly suitable for carbon K-edge NEXAFS. The endstation can be operated at pressures up to 100 mbar, whereby XPS can be routinely performed up to 30 mbar. A selection of typical data demonstrates the capability of the instrument to analyse details of the surface composition of solid samples under ambient-pressure conditions using XPS and NEXAFS. In addition, it offers a convenient way of analysing the gas phase through X-ray absorption spectroscopy. Short XPS spectra can be measured at a time scale of tens of seconds. The shortest data acquisition times for NEXAFS are around 0.5 s per data point.	Sep 2020
I15-1-X-ray Pair Distribution Function (XPDF)	Exploring the origins of crystallisation kinetics in hierarchical materials using in situ X-ray diffraction and pair distribution function analysis Matthew E. Potter , Mark E. Light , Daniel J. M. Irving , Alice E. Oakley , Stephanie Chapman , Philip Chater , Geoff Cutts , Andrew Watts , Michael Wharmby , Bart D. Vandegehuchte , Moritz W. Schreiber , Robert Raja Physical Chemistry Chemical Physics 336 DOI: 10.1039/D0CP00670J Diamond Proposal Number(s): [21878, 19039, 16135, 22842] Show Abstract ▼ Abstract: The discovery of novel catalytic materials is predicated on understanding contemporary synthetic processes. With this fundamental knowledge in place it becomes possible to modify the final material with subtle changes to the synthesis process. In this vein, hierarchical materials, formed by the addition of a mesoporogen within the hydrothermal synthesis, have attracted a significant amount of attention due to their catalytic benefits over analogous microporous species. In this work we monitor the hydrothermal synthesis in situ of a hierarchical and a microporous aluminophosphate, for the first time, combining total scattering and pairwise distribution function data. In doing so we observe the local formation of the species, and the longer range crystallisation processes concurrently.	Mar 2020

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