Search Results

You searched for all publications:

with publication states 'Published (Approved)'

Search Again...

These results only include publications that have been reviewed and processed by Diamond Light Source. To also view papers that have not yet been processed click here.

You can use the folder icon under Actions to collate papers as required. You can then click on View Folder in the left-hand navigation to review and/or download your folder.

Exact matches
Related results

5 items found (0 items not approved), displaying all items.1

Instruments / Technical Area	Publication Details	Published
I03-Macromolecular Crystallography I04-1-Macromolecular Crystallography (fixed wavelength) I04-Macromolecular Crystallography	A rotary mechanism for allostery in bacterial hybrid malic enzymes Christopher John Harding , Ian Thomas Cadby , Patrick Joseph Moynihan , Andrew Lee Lovering Nature Communications 12 DOI: 10.1038/s41467-021-21528-2 Diamond Proposal Number(s): [14692] Open Access Show Abstract ▼ Abstract: Bacterial hybrid malic enzymes (MaeB grouping, multidomain) catalyse the transformation of malate to pyruvate, and are a major contributor to cellular reducing power and carbon flux. Distinct from other malic enzyme subtypes, the hybrid enzymes are regulated by acetyl-CoA, a molecular indicator of the metabolic state of the cell. Here we solve the structure of a MaeB protein, which reveals hybrid enzymes use the appended phosphotransacetylase (PTA) domain to form a hexameric sensor that communicates acetyl-CoA occupancy to the malic enzyme active site, 60 Å away. We demonstrate that allostery is governed by a large-scale rearrangement that rotates the catalytic subunits 70° between the two states, identifying MaeB as a new model enzyme for the study of ligand-induced conformational change. Our work provides the mechanistic basis for metabolic control of hybrid malic enzymes, and identifies inhibition-insensitive variants that may find utility in synthetic biology.	Feb 2021
I03-Macromolecular Crystallography I04-Macromolecular Crystallography	Bypassing the requirement for aminoacyl-tRNA by a cyclodipeptide synthase enzyme Christopher Harding , Emmajay Sutherland , Jane G. Hanna , Douglas R. Houston , Clarissa M. Czekster Rsc Chemical Biology 9 DOI: 10.1039/D0CB00142B Diamond Proposal Number(s): [19844] Open Access Show Abstract ▼ Abstract: Cyclodipeptide synthases (CDPSs) produce a variety of cyclic dipeptide products by utilising two aminoacylated tRNA substrates. We sought to investigate the minimal requirements for substrate usage in this class of enzymes as the relationship between CDPSs and their substrates remains elusive. Here, we investigated the Bacillus thermoamylovorans enzyme, BtCDPS, which synthesises cyclo(L-Leu–L-Leu). We systematically tested where specificity arises and, in the process, uncovered small molecules (activated amino esters) that will suffice as substrates, although catalytically poor. We solved the structure of BtCDPS to 1.7 Å and combining crystallography, enzymatic assays and substrate docking experiments propose a model for how the minimal substrates interact with the enzyme. This work is the first report of a CDPS enzyme utilizing a molecule other than aa-tRNA as a substrate; providing insights into substrate requirements and setting the stage for the design of improved simpler substrates.	Jan 2021
I03-Macromolecular Crystallography I04-1-Macromolecular Crystallography (fixed wavelength) I04-Macromolecular Crystallography	A lysozyme with altered substrate specificity facilitates prey cell exit by the periplasmic predator Bdellovibrio bacteriovorus Christopher J. Harding , Simona G. Huwiler , Hannah Somers , Carey Lambert , Luke J. Ray , Rob Till , Georgina Taylor , Patrick J. Moynihan , R. Elizabeth Sockett , Andrew L. Lovering Nature Communications 11 DOI: 10.1038/s41467-020-18139-8 Diamond Proposal Number(s): [14692, 19880] Open Access Show Abstract ▼ Abstract: Lysozymes are among the best-characterized enzymes, acting upon the cell wall substrate peptidoglycan. Here, examining the invasive bacterial periplasmic predator Bdellovibrio bacteriovorus, we report a diversified lysozyme, DslA, which acts, unusually, upon (GlcNAc-) deacetylated peptidoglycan. B. bacteriovorus are known to deacetylate the peptidoglycan of the prey bacterium, generating an important chemical difference between prey and self walls and implying usage of a putative deacetyl-specific “exit enzyme”. DslA performs this role, and ΔDslA strains exhibit a delay in leaving from prey. The structure of DslA reveals a modified lysozyme superfamily fold, with several adaptations. Biochemical assays confirm DslA specificity for deacetylated cell wall, and usage of two glutamate residues for catalysis. Exogenous DslA, added ex vivo, is able to prematurely liberate B. bacteriovorus from prey, part-way through the predatory lifecycle. We define a mechanism for specificity that invokes steric selection, and use the resultant motif to identify wider DslA homologues.	Sep 2020
I02-Macromolecular Crystallography I03-Macromolecular Crystallography I04-1-Macromolecular Crystallography (fixed wavelength) I04-Macromolecular Crystallography	Target highlights in CASP13: Experimental target structures through the eyes of their authors Rosalba Lepore , Andriy Kryshtafovych , Markus Alahuhta , Harshul A. Veraszto , Yannick J. Bomble , Joshua C. Bufton , Alex N. Bullock , Cody Caba , Hongnan Cao , Owen R. Davies , Ambroise Desfosses , Matthew Dunne , Krzysztof Fidelis , Celia W. Goulding , Manickam Gurusaran , Irina Gutsche , Christopher J. Harding , Marcus D. Hartmann , Christopher S. Hayes , Andrzej Joachimiak , Petr G. Leiman , Peter Loppnau , Andrew L. Lovering , Vladimir V. Lunin , Karolina Michalska , Ignacio Mir‐sanchis , Alok Mitra , John Moult , George N. Phillips Jr , Daniel Pinkas , Phoebe A. Rice , Yufeng Tong , Maya Topf , Jonathan D. Walton , Torsten Schwede Proteins: Structure, Function, And Bioinformatics 7 DOI: 10.1002/prot.25805 Diamond Proposal Number(s): [14692] Open Access Show Abstract ▼ Abstract: The functional and biological significance of selected CASP13 targets are described by the authors of the structures. The structural biologists discuss the most interesting structural features of the target proteins and assess whether these features were correctly reproduced in the predictions submitted to the CASP13 experiment.	Sep 2019
I04-Macromolecular Crystallography	Evidence for phospholipid export from the bacterial inner membrane by the Mla ABC transport system Gareth W. Hughes , Stephen C. L. Hall , Claire S. Laxton , Pooja Sridhar , Amirul H. Mahadi , Caitlin Hatton , Thomas J. Piggot , Peter J. Wotherspoon , Aneika C. Leney , Douglas G. Ward , Mohammed Jamshad , Vaclav Spana , Ian T. Cadby , Christopher Harding , Georgia L. Isom , Jack A. Bryant , Rebecca J. Parr , Yasin Yakub , Mark Jeeves , Damon Huber , Ian R. Henderson , Luke A. Clifton , Andrew L. Lovering , Timothy J. Knowles Nature Microbiology 106 DOI: 10.1038/s41564-019-0481-y Diamond Proposal Number(s): [14692] Show Abstract ▼ Abstract: The Mla pathway is believed to be involved in maintaining the asymmetrical Gram-negative outer membrane via retrograde phospholipid transport. The pathway is composed of three components: the outer membrane MlaA–OmpC/F complex, a soluble periplasmic protein, MlaC, and the inner membrane ATPase, MlaFEDB complex. Here, we solve the crystal structure of MlaC in its phospholipid-free closed apo conformation, revealing a pivoting β-sheet mechanism that functions to open and close the phospholipid-binding pocket. Using the apo form of MlaC, we provide evidence that the inner-membrane MlaFEDB machinery exports phospholipids to MlaC in the periplasm. Furthermore, we confirm that the phospholipid export process occurs through the MlaD component of the MlaFEDB complex and that this process is independent of ATP. Our data provide evidence of an apparatus for lipid export away from the inner membrane and suggest that the Mla pathway may have a role in anterograde phospholipid transport.	Jun 2019

Publications Database

Search Results

You searched for all publications:

Search Again...

Subject Areas (check all that apply) select all

Instruments used to collect data (check all that apply) select all

Collaborations involved (check all that apply) select all

Diamond Offline Facilities used

Relevant Technical Areas (check all that apply) select all

Menu for Anonymous User