I19-Small Molecule Single Crystal Diffraction
|
Diamond Proposal Number(s):
[20876]
Abstract: A new metastable phase in flash-frozen disordered Prussian blue analogues is reported. The phase is characterised by the appearance of diffuse scattering clouds and the reduction of the local structure symmetry: from cubic to a tetragonal or lower space group. The phase transition is characterised by the translational modulation of the structure and is likely caused by the freezing of the water confined in the pores of the structure.
|
Jun 2022
|
|
I24-Microfocus Macromolecular Crystallography
|
Chih-Jen
Wei
,
Wei
Bu
,
Laura A.
Nguyen
,
Joseph D.
Batchelor
,
Junghyun
Kim
,
Stefania
Pittaluga
,
James R.
Fuller
,
Hanh
Nguyen
,
Te-Hui
Chou
,
Jeffrey I.
Cohen
,
Gary J.
Nabel
Diamond Proposal Number(s):
[22894]
Abstract: Epstein-Barr virus (EBV) is the major cause of infectious mononucleosis and is associated with several human cancers and, more recently, multiple sclerosis. Despite its prevalence and health impact, there are currently no vaccines or treatments. Four viral glycoproteins (gp), gp350 and gH/gL/gp42, mediate entry into the major sites of viral replication, B cells, and epithelial cells. Here, we designed a nanoparticle vaccine displaying these proteins and showed that it elicits potent neutralizing antibodies that protect against infection in vivo. We designed single-chain gH/gL and gH/gL/gp42 proteins that were each fused to bacterial ferritin to form a self-assembling nanoparticle. Structural analysis revealed that single-chain gH/gL and gH/gL/gp42 adopted a similar conformation to the wild-type proteins, and the protein spikes were observed by electron microscopy. Single-chain gH/gL or gH/gL/gp42 nanoparticle vaccines were constructed to ensure product homogeneity needed for clinical development. These vaccines elicited neutralizing antibodies in mice, ferrets, and nonhuman primates that inhibited EBV entry into both B cells and epithelial cells. When mixed with a previously reported gp350 nanoparticle vaccine, gp350D123, no immune competition was observed. To confirm its efficacy in vivo, humanized mice were challenged with EBV after passive transfer of IgG from mice vaccinated with control, gH/gL/gp42+gp350D123, or gH/gL+gp350D123 nanoparticles. Although all control animals were infected, only one mouse in each vaccine group that received immune IgG had detectable transient viremia. Furthermore, no EBV lymphomas were detected in immune animals. This bivalent EBV nanoparticle vaccine represents a promising candidate to prevent EBV infection and EBV-related malignancies in humans.
|
May 2022
|
|
B07-C-Versatile Soft X-ray beamline: Ambient Pressure XPS and NEXAFS
|
Diamond Proposal Number(s):
[25766]
Open Access
Abstract: Conductometric gas sensors (CGS) provide a reproducible gas response at a low cost but their operation mechanisms are still not fully understood. In this paper, we elucidate the nature of interactions between SnO2, a common gas-sensitive material, and O2, a ubiquitous gas central to the detection mechanisms of CGS. Using synchrotron radiation, we investigated a working SnO2 sensor under operando conditions via near-ambient pressure (NAP) XPS with simultaneous resistance measurements, and created a depth profile of the variable near-surface stoichiometry of SnO2−x as a function of O2 pressure. Our results reveal a correlation between the dynamically changing surface oxygen vacancies and the resistance response in SnO2-based CGS. While oxygen adsorbates were observed in this study we conclude that these are an intermediary in oxygen transport between the gas phase and the lattice, and that surface oxygen vacancies, not the observed oxygen adsorbates, are central to response generation in SnO2-based gas sensors.
|
May 2022
|
|
B21-High Throughput SAXS
I04-Macromolecular Crystallography
|
Diamond Proposal Number(s):
[14739, 29790, 20229]
Open Access
Abstract: IMP dehydrogenase(IMPDH) is an essential enzyme that catalyzes the rate-limiting step in the guanine nucleotide pathway. In eukaryotic cells, GTP binding to the regulatory domain allosterically controls the activity of IMPDH by a mechanism that is fine-tuned by post-translational modifications and enzyme polymerization. Nonetheless, the mechanisms of regulation of IMPDH in bacterial cells remain unclear. Using biochemical, structural, and evolutionary analyses, we demonstrate that, in most bacterial phyla, (p)ppGpp compete with ATP to allosterically modulate IMPDH activity by binding to a, previously unrecognized, conserved high affinity pocket within the regulatory domain. This pocket was lost during the evolution of Proteobacteria, making their IMPDHs insensitive to these alarmones. Instead, most proteobacterial IMPDHs evolved to be directly modulated by the balance between ATP and GTP that compete for the same allosteric binding site. Altogether, we demonstrate that the activity of bacterial IMPDHs is allosterically modulated by a universally conserved nucleotide-controlled conformational switch that has divergently evolved to adapt to the specific particularities of each organism. These results reconcile the reported data on the crosstalk between (p)ppGpp signaling and the guanine nucleotide biosynthetic pathway and reinforce the essential role of IMPDH allosteric regulation on bacterial GTP homeostasis.
|
May 2022
|
|
I08-Scanning X-ray Microscopy beamline (SXM)
|
Dawn M.
Buchanan
,
Laura
Newsome
,
Jonathan R.
Lloyd
,
Majid
Kazemian
,
Burkhard
Kaulich
,
Tohru
Araki
,
Heath
Bagshaw
,
John
Waters
,
Gerrit
Van Der Laan
,
Alpha
N’diaye
,
Victoria S.
Coker
Diamond Proposal Number(s):
[17626]
Open Access
Abstract: Cobalt is an essential element for life and plays a crucial role in supporting the drive to clean energy, due to its importance in rechargeable batteries. Co is often associated with Fe in the environment, but the fate of Co in Fe-rich biogeochemically-active environments is poorly understood. To address this, synchrotron-based scanning X-ray microscopy (SXM) was used investigate the behaviour of cobalt at the nanoscale in Co-Fe(III)-oxyhydroxides undergoing microbial reduction. SXM can assess spatial changes in metal speciation and organic compounds helping to elucidate the electron transfer processes occurring at the cell-mineral interface and inform on the fate of cobalt in redox horizons. G. sulfurreducens was used to reduce synthetic Co-ferrihydrite as an analogue of natural cobalt-iron-oxides. Magnetite [Fe(II)/Fe(III)3O4] production was confirmed by powder X-ray diffraction (XRD), SXM and X-ray magnetic circular dichroism (XMCD) data, where best fits of the latter suggested Co-bearing magnetite. Macro-scale XAS techniques suggested Co(III) reduction occurred and complementary SXM at the nanoscale, coupled with imaging, found localised biogenic Co(III) reduction at the cell-mineral interface via direct contact with outer membrane cytochromes. No discernible localised changes in Fe speciation were detected in the reordered cobalt-iron-oxides that were formed and at the end point of the experiment only 11% Co and 1.5% Fe had been solubilised. The solid phase retention, alongside the highly localised and preferential cobalt bioreduction observed at the nanoscale is consistent with retention of Co in redox zones. This work improves our fundamental molecular-scale understanding of the fate of Co in complex environmental systems and supports the development of biogenic Co-doped magnetite for industrial applications from drug delivery systems to magnetic recording media.
|
May 2022
|
|
I19-Small Molecule Single Crystal Diffraction
|
Lei
Ji
,
Stefan
Riese
,
Alexander
Schmiedel
,
Marco
Holzapfel
,
Maximillian
Fest
,
Jörn
Nitsch
,
Basile F. E.
Curchod
,
Alexandra
Friedrich
,
Lin
Wu
,
Hamad H.
Al Mamari
,
Sebastian
Hammer
,
Jens
Pflaum
,
Mark A.
Fox
,
David J.
Tozer
,
Maik
Finze
,
Christoph
Lambert
,
Todd B.
Marder
Diamond Proposal Number(s):
[18456]
Open Access
Abstract: Reversible conversion between excited-states plays an important role in many photophysical phenomena. Using 1-(pyren-2′-yl)-o-carborane as a model, we studied the photoinduced reversible charge-transfer (CT) process and the thermodynamic equilibrium between the locally-excited (LE) state and CT state, by combining steady state, time-resolved, and temperature-dependent fluorescence spectroscopy, fs- and ns-transient absorption, and DFT and LR-TDDFT calculations. Our results show that the energy gaps and energy barriers between the LE, CT, and a non-emissive ‘mixed’ state of 1-(pyren-2′-yl)-o-carborane are very small, and all three excited states are accessible at room temperature. The internal-conversion and reverse internal-conversion between LE and CT states are significantly faster than the radiative decay, and the two states have the same lifetimes and are in thermodynamic equilibrium.
|
Apr 2022
|
|
I06-Nanoscience
|
Filip
Krizek
,
Sonka
Reimers
,
Zdeněk
Kašpar
,
Alberto
Marmodoro
,
Jan
Michalička
,
Ondřej
Man
,
Alexander
Edström
,
Oliver J.
Amin
,
Kevin W.
Edmonds
,
Richard P.
Campion
,
Francesco
Maccherozzi
,
Sarnjeet S.
Dhesi
,
Jan
Zubáč
,
Domink
Kriegner
,
Dina
Carbone
,
Jakub
Železný
,
Karel
Výborný
,
Kamil
Olejník
,
Vít
Novák
,
Jan
Rusz
,
Juan-Carlos
Idrobo
,
Peter
Wadley
,
Tomas
Jungwirth
Diamond Proposal Number(s):
[22437]
Open Access
Abstract: The interest in understanding scaling limits of magnetic textures such as domain walls spans the entire field of magnetism from its physical fundamentals to applications in information technologies. Here, we explore antiferromagnetic CuMnAs in which imaging by x-ray photoemission reveals the presence of magnetic textures down to nanoscale, reaching the detection limit of this established microscopy in antiferromagnets. We achieve atomic resolution by using differential phase-contrast imaging within aberration-corrected scanning transmission electron microscopy. We identify abrupt domain walls in the antiferromagnetic film corresponding to the Néel order reversal between two neighboring atomic planes. Our work stimulates research of magnetic textures at the ultimate atomic scale and sheds light on electrical and ultrafast optical antiferromagnetic devices with magnetic field–insensitive neuromorphic functionalities.
|
Apr 2022
|
|
I20-Scanning-X-ray spectroscopy (XAS/XES)
|
Diamond Proposal Number(s):
[21441]
Open Access
Abstract: Selenium (Se) is a toxic contaminant with multiple anthropogenic sources, including 79Se from nuclear fission. Se mobility in the geosphere is generally governed by its oxidation state, therefore understanding Se speciation under variable redox conditions is important for the safe management of Se contaminated sites. Here, we investigate Se behavior in sediment groundwater column systems. Experiments were conducted with environmentally relevant Se concentrations, using a range of groundwater compositions, and the impact of electron-donor (i.e., biostimulation) and groundwater sulfate addition was examined over a period of 170 days. X-Ray Absorption Spectroscopy and standard geochemical techniques were used to track changes in sediment associated Se concentration and speciation. Electron-donor amended systems with and without added sulfate retained up to 90% of added Se(VI)(aq), with sediment associated Se speciation dominated by trigonal Se(0) and possibly trace Se(-II); no Se colloid formation was observed. The remobilization potential of the sediment associated Se species was then tested in reoxidation and seawater intrusion perturbation experiments. In all treatments, sediment associated Se (i.e., trigonal Se(0)) was largely resistant to remobilization over the timescales of the experiments (170 days). However, in the perturbation experiments, less Se was remobilized from sulfidic sediments, suggesting that previous sulfate-reducing conditions may buffer Se against remobilization and migration.
|
Apr 2022
|
|
I03-Macromolecular Crystallography
I04-Macromolecular Crystallography
I24-Microfocus Macromolecular Crystallography
|
Nathan R.
Zaccai
,
Zuzana
Kadlecova
,
Veronica
Kane Dickson
,
Kseniya
Korobchevskaya
,
Jan
Kamenicky
,
Oleksiy
Kovtun
,
Perunthottathu K.
Umasankar
,
Antoni G.
Wrobel
,
Jonathan G. G.
Kaufman
,
Sally R.
Gray
,
Kun
Qu
,
Philip R.
Evans
,
Marco
Fritzsche
,
Filip
Sroubek
,
Stefan
Höning
,
John A. G.
Briggs
,
Bernard T.
Kelly
,
David J.
Owen
,
Linton M.
Traub
Abstract: Clathrin-mediated endocytosis (CME) is the main mechanism by which mammalian cells control their cell surface proteome. Proper operation of the pivotal CME cargo adaptor AP2 requires membrane-localized Fer/Cip4 homology domain-only proteins (FCHO). Here, live-cell enhanced total internal reflection fluorescence–structured illumination microscopy shows that FCHO marks sites of clathrin-coated pit (CCP) initiation, which mature into uniform-sized CCPs comprising a central patch of AP2 and clathrin corralled by an FCHO/Epidermal growth factor potential receptor substrate number 15 (Eps15) ring. We dissect the network of interactions between the FCHO interdomain linker and AP2, which concentrates, orients, tethers, and partially destabilizes closed AP2 at the plasma membrane. AP2’s subsequent membrane deposition drives its opening, which triggers FCHO displacement through steric competition with phosphatidylinositol 4,5-bisphosphate, clathrin, cargo, and CME accessory factors. FCHO can now relocate toward a CCP’s outer edge to engage and activate further AP2s to drive CCP growth/maturation.
|
Apr 2022
|
|
Krios I-Titan Krios I at Diamond
|
Abstract: Kinetochores assemble onto specialized centromeric CENP-A nucleosomes (CENP-ANuc) to mediate attachments between chromosomes and the mitotic spindle. We describe cryo-EM structures of the human inner kinetochore CCAN (Constitutive Centromere Associated Network) complex bound to CENP-ANuc reconstituted onto α-satellite DNA. CCAN forms edge-on contacts with CENP-ANuc, while a linker DNA segment of the α-satellite repeat emerges from the fully-wrapped end of the nucleosome to thread through the central CENP-LN channel that tightly grips the DNA. The CENP-TWSX histone-fold module further augments DNA binding and partially wraps the linker DNA in a manner reminiscent of canonical nucleosomes. Our study suggests that the topological entrapment of the linker DNA by CCAN provides a robust mechanism by which kinetochores withstand both pushing and pulling forces exerted by the mitotic spindle.
|
Apr 2022
|
|
