Krios I-Titan Krios I at Diamond
Krios II-Titan Krios II at Diamond
Krios IV-Titan Krios IV at Diamond
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Jianbing
Ma
,
Gangshun
Yi
,
Mingda
Ye
,
Craig
Macgregor-Chatwin
,
Yuewen
Sheng
,
Ying
Lu
,
Ming
Li
,
Qingrong
Li
,
Dong
Wang
,
Robert J. C.
Gilbert
,
Peijun
Zhang
Diamond Proposal Number(s):
[29812]
Open Access
Abstract: The cryo-electron microscopy (cryoEM) method has enabled high-resolution structure determination of numerous biomolecules and complexes. Nevertheless, cryoEM sample preparation of challenging proteins and complexes, especially those with low abundance or with preferential orientation, remains a major hurdle. We developed an affinity-grid method employing monodispersed single particle streptavidin on a lipid monolayer to enhance particle absorption on the grid surface and alleviate sample exposure to the air-water interface. Using this approach, we successfully enriched the Thermococcus kodakarensis mini-chromosome maintenance complex 3 (MCM3) on cryoEM grids through biotinylation and resolved its structure. We further utilized this affinity method to tether the biotin-tagged dsDNA to selectively enrich a stable MCM3-ATP-dsDNA complex for cryoEM structure determination. Intriguingly, both MCM3 apo and dsDNA bound structures exhibit left-handed open spiral conformations, distinct from other reported MCM structures. The large open gate is sufficient to accommodate a dsDNA which could potentially be melted. The value of mspSA affinity method was further demonstrated by mitigating the issue of preferential angular distribution of HIV-1 capsid protein hexamer and RNA polymerase II elongation complex from Saccharomyces cerevisiae.
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Dec 2024
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B18-Core EXAFS
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Jia-Wei
Wang
,
Fengyi
Zhao
,
Lucia
Velasco
,
Maxime
Sauvan
,
Dooshaye
Moonshiram
,
Martina
Salati
,
Zhi-Mei
Luo
,
Sheng
He
,
Tao
Jin
,
Yan-Fei
Mu
,
Mehmed Z.
Ertem
,
Tianquan
Lian
,
Antoni
Llobet
Diamond Proposal Number(s):
[33134]
Open Access
Abstract: The selective photoreduction of CO2 in aqueous media based on earth-abundant elements only, is today a challenging topic. Here we present the anchoring of discrete molecular catalysts on organic polymeric semiconductors via covalent bonding, generating molecular hybrid materials with well-defined active sites for CO2 photoreduction, exclusively to CO in purely aqueous media. The molecular catalysts are based on aryl substituted Co phthalocyanines that can be coordinated by dangling pyridyl attached to a polymeric covalent triazine framework that acts as a light absorber. This generates a molecular hybrid material that efficiently and selectively achieves the photoreduction of CO2 to CO in KHCO3 aqueous buffer, giving high yields in the range of 22 mmol g−1 (458 μmol g−1 h−1) and turnover numbers above 550 in 48 h, with no deactivation and no detectable H2. The electron transfer mechanism for the activation of the catalyst is proposed based on the combined results from time-resolved fluorescence spectroscopy, in situ spectroscopies and quantum chemical calculations.
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Nov 2024
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I21-Resonant Inelastic X-ray Scattering (RIXS)
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Xiaoyang
Chen
,
Jaewon
Choi
,
Zhicheng
Jiang
,
Jiong
Mei
,
Kun
Jiang
,
Jie
Li
,
Stefano
Agrestini
,
Mirian
Garcia-Fernandez
,
Hualei
Sun
,
Xing
Huang
,
Dawei
Shen
,
Meng
Wang
,
Jiangping
Hu
,
Yi
Lu
,
Ke-Jin
Zhou
,
Donglai
Feng
Diamond Proposal Number(s):
[35805]
Open Access
Abstract: High-temperature superconductivity was discovered in the pressurized nickelate La3Ni2O7 which has a unique bilayer structure and mixed valence state of nickel. The properties at ambient pressure contain crucial information of the fundamental interactions and bosons mediating superconducting pairing. Here, using X-ray absorption spectroscopy and resonant inelastic X-ray scattering, we identified that Ni 3, Ni 3, and ligand oxygen 2p orbitals dominate the low-energy physics with a small charge-transfer energy. Well-defined optical-like magnetic excitations soften into quasi-static spin-density-wave ordering, evidencing the strong electronic correlation and rich magnetic properties. Based on an effective Heisenberg spin model, we extract a much stronger inter-layer effective magnetic superexchange than the intra-layer ones and propose two viable magnetic structures. Our findings emphasize that the Ni 3 orbital bonding within the bilayer induces novel electronic and magnetic excitations, setting the stage for further exploration of La3Ni2O7 superconductor.
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Nov 2024
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I11-High Resolution Powder Diffraction
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Diamond Proposal Number(s):
[31308]
Open Access
Abstract: Magnetic materials are composed of the simple building blocks of magnetic moments on a crystal lattice that interact via magnetic exchange. Yet from this simplicity emerges a remarkable diversity of magnetic states. Some reveal the deep quantum mechanical origins of magnetism, for example, quantum spin liquid (QSL) states in which magnetic moments remain disordered at low temperatures despite being strongly correlated through quantum entanglement. A promising theoretical model of a QSL is the Kitaev model, composed of unusual bond-dependent exchange interactions, but experimentally, this model is challenging to realise. Here we show that the material requirements for the Kitaev QSL survive an extended pseudo-edge-sharing superexchange pathway of Ru3+ octahedra within the honeycomb layers of the inorganic framework solid, RuP3SiO11. We confirm the requisite state of Ru3+ in RuP3SiO11 and resolve the hierarchy of exchange interactions that provide experimental access to an unexplored region of the Kitaev model.
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Nov 2024
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I04-Macromolecular Crystallography
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Gavuthami
Murugesan
,
Rachel L.
Paterson
,
Rakesh
Kulkarni
,
Veronica
Ilkow
,
Richard J.
Suckling
,
Mary M.
Connolly
,
Vijaykumar
Karuppiah
,
Robert
Pengelly
,
Archana
Jadhav
,
Jose
Donoso
,
Tiaan
Heunis
,
Wilawan
Bunjobpol
,
Gwilym
Philips
,
Kafayat
Ololade
,
Daniel
Kay
,
Anshuk
Sarkar
,
Claire
Barber
,
Ritu
Raj
,
Carole
Perot
,
Tressan
Grant
,
Agatha
Treveil
,
Andrew
Walker
,
Marcin
Dembek
,
Dawn
Gibbs-Howe
,
Miriam
Hock
,
Ricardo J.
Carreira
,
Kate E.
Atkin
,
Lucy
Dorrell
,
Andrew
Knox
,
Sarah
Leonard
,
Mariolina
Salio
,
Luis F.
Godinho
Diamond Proposal Number(s):
[28224]
Open Access
Abstract: The non-polymorphic HLA-E molecule offers opportunities for new universal immunotherapeutic approaches to chronic infectious diseases. Chronic Hepatitis B virus (HBV) infection is driven in part by T cell dysfunction due to elevated levels of the HBV envelope (Env) protein hepatitis B surface antigen (HBsAg). Here we report the characterization of three genotypic variants of an HLA-E-binding HBsAg peptide, Env371-379, identified through bioinformatic predictions and verified by biochemical and cellular assays. Using a soluble affinity-enhanced T cell receptor (TCR) (a09b08)-anti-CD3 bispecific molecule to probe HLA-E presentation of the Env371-379 peptides, we demonstrate that only the most stable Env371-379 variant, L6I, elicits functional responses to a09b08-anti-CD3-redirected polyclonal T cells co-cultured with targets expressing endogenous HBsAg. Furthermore, HLA-E-Env371-379 L6I-specific CD8+ T cells are detectable in HBV-naïve donors and people with chronic HBV after in vitro priming. In conclusion, we provide evidence for HLA-E-mediated HBV Env peptide presentation, and highlight the effect of viral mutations on the stability and targetability of pHLA-E molecules.
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Nov 2024
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I04-1-Macromolecular Crystallography (fixed wavelength)
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Martin P.
Schwalm
,
Johannes
Dopfer
,
Adarsh
Kumar
,
Francesco A.
Greco
,
Nicolas
Bauer
,
Frank
Löhr
,
Jan
Heering
,
Sara
Cano-Franco
,
Severin
Lechner
,
Thomas
Hanke
,
Ivana
Jaser
,
Viktoria
Morasch
,
Christopher
Lenz
,
Daren
Fearon
,
Peter G.
Marples
,
Charles W. E.
Tomlinson
,
Lorene
Brunello
,
Krishna
Saxena
,
Nathan B. P.
Adams
,
Frank
Von Delft
,
Susanne
Müller
,
Alexandra
Stolz
,
Ewgenij
Proschak
,
Bernhard
Kuster
,
Stefan
Knapp
,
Vladimir V.
Rogov
Diamond Proposal Number(s):
[29658]
Open Access
Abstract: Recent successes in developing small molecule degraders that act through the ubiquitin system have spurred efforts to extend this technology to other mechanisms, including the autophagosomal-lysosomal pathway. Therefore, reports of autophagosome tethering compounds (ATTECs) have received considerable attention from the drug development community. ATTECs are based on the recruitment of targets to LC3/GABARAP, a family of ubiquitin-like proteins that presumably bind to the autophagosome membrane and tether cargo-loaded autophagy receptors into the autophagosome. In this work, we rigorously tested the target engagement of the reported ATTECs to validate the existing LC3/GABARAP ligands. Surprisingly, we were unable to detect interaction with their designated target LC3 using a diversity of biophysical methods. Intrigued by the idea of developing ATTECs, we evaluated the ligandability of LC3/GABARAP by in silico docking and large-scale crystallographic fragment screening. Data based on approximately 1000 crystal structures revealed that most fragments bound to the HP2 but not to the HP1 pocket within the LIR docking site, suggesting a favorable ligandability of HP2. Through this study, we identified diverse validated LC3/GABARAP ligands and fragments as starting points for chemical probe and ATTEC development.
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Nov 2024
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I07-Surface & interface diffraction
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Benjamin M.
Gallant
,
Philippe
Holzhey
,
Joel A.
Smith
,
Saqlain
Choudhary
,
Karim A.
Elmestekawy
,
Pietro
Caprioglio
,
Igal
Levine
,
Alexandra A.
Sheader
,
Esther Y.-H.
Hung
,
Fengning
Yang
,
Daniel T. W.
Toolan
,
Rachel C.
Kilbride
,
Karl-Augustin
Zaininger
,
James M.
Ball
,
M. Greyson
Christoforo
,
Nakita K.
Noel
,
Laura M.
Herz
,
Dominik J.
Kubicki
,
Henry J.
Snaith
Diamond Proposal Number(s):
[33462]
Open Access
Abstract: Perovskite solar cells (PSCs) offer an efficient, inexpensive alternative to current photovoltaic technologies, with the potential for manufacture via high-throughput coating methods. However, challenges for commercial-scale solution-processing of metal-halide perovskites include the use of harmful solvents, the expense of maintaining controlled atmospheric conditions, and the inherent instabilities of PSCs under operation. Here, we address these challenges by introducing a high volatility, low toxicity, biorenewable solvent system to fabricate a range of 2D perovskites, which we use as highly effective precursor phases for subsequent transformation to α-formamidinium lead triiodide (α-FAPbI3), fully processed under ambient conditions. PSCs utilising our α-FAPbI3 reproducibly show remarkable stability under illumination and elevated temperature (ISOS-L-2) and “damp heat” (ISOS-D-3) stressing, surpassing other state-of-the-art perovskite compositions. We determine that this enhancement is a consequence of the 2D precursor phase crystallisation route, which simultaneously avoids retention of residual low-volatility solvents (such as DMF and DMSO) and reduces the rate of degradation of FA+ in the material. Our findings highlight both the critical role of the initial crystallisation process in determining the operational stability of perovskite materials, and that neat FA+-based perovskites can be competitively stable despite the inherent metastability of the α-phase.
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Nov 2024
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B21-High Throughput SAXS
I04-Macromolecular Crystallography
I24-Microfocus Macromolecular Crystallography
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Alena
Kroupova
,
Valentina A.
Spiteri
,
Zoe J.
Rutter
,
Hirotake
Furihata
,
Darren
Darren
,
Sarath
Ramachandran
,
Sohini
Chakraborti
,
Kevin
Haubrich
,
Julie
Pethe
,
Denzel
Gonzales
,
Andre J.
Wijaya
,
Maria
Rodriguez-Rios
,
Manon
Sturbaut
,
Dylan M.
Lynch
,
William
Farnaby
,
Mark A.
Nakasone
,
David
Zollman
,
Alessio
Ciulli
Diamond Proposal Number(s):
[26793, 35324, 33832, 38813]
Open Access
Abstract: The ubiquitin E3 ligase cereblon (CRBN) is the target of therapeutic drugs thalidomide and lenalidomide and is recruited by most targeted protein degraders (PROTACs and molecular glues) in clinical development. Biophysical and structural investigation of CRBN has been limited by current constructs that either require co-expression with the adaptor DDB1 or inadequately represent full-length protein, with high-resolution structures of degrader ternary complexes remaining rare. We present the design of CRBNmidi, a construct that readily expresses from E. coli with high yields as soluble, stable protein without DDB1. We benchmark CRBNmidi for wild-type functionality through a suite of biophysical techniques and solve high-resolution co-crystal structures of its binary and ternary complexes with degraders. We qualify CRBNmidi as an enabling tool to accelerate structure-based discovery of the next generation of CRBN based therapeutics.
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Oct 2024
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I06-Nanoscience (XPEEM)
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Alessandra
Milloch
,
Ignacio
Figueruelo-Campanero
,
Wei-Fan
Hsu
,
Selene
Mor
,
Simon
Mellaerts
,
Francesco
Maccherozzi
,
Larissa S. I.
Veiga
,
Sarnjeet S.
Dhesi
,
Mauro
Spera
,
Jin Won
Seo
,
Jean-Pierre
Locquet
,
Michele
Fabrizio
,
Mariela
Menghini
,
Claudio
Giannetti
Diamond Proposal Number(s):
[27218, 31711, 34455]
Open Access
Abstract: Avalanche resistive switching is the fundamental process that triggers the sudden change of the electrical properties in solid-state devices under the action of intense electric fields. Despite its relevance for information processing, ultrafast electronics, neuromorphic devices, resistive memories and brain-inspired computation, the nature of the local stochastic fluctuations that drive the formation of metallic regions within the insulating state has remained hidden. Here, using operando X-ray nano-imaging, we have captured the origin of resistive switching in a V2O3-based device under working conditions. V2O3 is a paradigmatic Mott material, which undergoes a first-order metal-to-insulator phase transition together with a lattice transformation that breaks the threefold rotational symmetry of the rhombohedral metallic phase. We reveal a new class of volatile electronic switching triggered by nanoscale topological defects appearing in the shear-strain based order parameter that describes the insulating phase. Our results pave the way to the use of strain engineering approaches to manipulate such topological defects and achieve the full dynamical control of the electronic Mott switching. Topology-driven, reversible electronic transitions are relevant across a broad range of quantum materials, comprising transition metal oxides, chalcogenides and kagome metals.
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Oct 2024
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Krios IV-Titan Krios IV at Diamond
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Diamond Proposal Number(s):
[23047]
Open Access
Abstract: Respiratory complex I is pivotal for cellular energy conversion, harnessing energy from NADH:ubiquinone oxidoreduction to drive protons across energy-transducing membranes for ATP synthesis. Despite detailed structural information on complex I, its mechanism of catalysis remains elusive due to lack of accompanying functional data for comprehensive structure-function analyses. Here, we present the 2.3-Å resolution structure of complex I from the α-proteobacterium Paracoccus denitrificans, a close relative of the mitochondrial progenitor, in phospholipid-bilayer nanodiscs. Three eukaryotic-type supernumerary subunits (NDUFS4, NDUFS6 and NDUFA12) plus a novel L-isoaspartyl-O-methyltransferase are bound to the core complex. Importantly, the enzyme is in a single, homogeneous resting state that matches the closed, turnover-ready (active) state of mammalian complex I. Our structure reveals the elements that stabilise the closed state and completes P. denitrificans complex I as a unified platform for combining structure, function and genetics in mechanistic studies.
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Oct 2024
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