Diamond Annual Review 2023/24

56 D I A M O N D L I G H T S O U R C E A N N U A L R E V I E W 2 0 2 3 / 2 4 Integrated Facilities and Collaborations A s a world-leading centre for synchrotron science and a cornerstone of a world-class site for scientific discovery and innovation at Harwell, Diamond Light Source has powerful synergies with its neighbouring research institutes and beyond the campus, through collaborations and shared visions. The integrated facilities at Diamond present academic and industrial users with a one-stop-shop for research opportunities, enabling them to combine cutting-edge techniques and capabilities to advance their studies. During the period 2023/24, Diamond was active on over 72grant fundedprojects (16ofwhichwereDiamond led). Our grant portfolioholds projectswithbothnational and international collaborators, where Diamond has contributed to projects worth over £248m. The Membrane Protein Lab TheMembrane Protein Laboratory (MPL), at Diamond is aWellcome funded resource that supports integratedmembrane protein structural biology. Located within the Research Complex at Harwell, the MPL enables membranes protein research through the delivery of high-quality samples to Diamond’s beamlines and microscopes as well as providing a platform to support membrane protein biochemistry. Membrane proteins are found at the junctions between the outside world and the inner workings of the cell. Multicellular organisms such as humans use membrane proteins for communication, to acquire nutrients and detect threats. Membrane proteins are important targets for biomedicine and biotechnology. Over half of all medicines alter membrane protein function. Understanding the structure and function of these proteins in isolation as well as within the wider cellular context will helps us to develop new therapeutics to tackle disease. Researchers from the University of Oxford and the Rosalind Franklin Institute were able to solve the structure of a bacterial membrane protein complex that consists of an oligosaccharide polymeraseWzy and its regulating co-polymerase, Wzz [1]. The structure of Wzy:Wzz complex alongside in vivo experiments show how the polymerisation of Wzy is regulated by Wzz giving a new structural insight into a conserved mechanism that regulates polysaccharide chain length in bacteria. In other work, research led by a University of Nottingham and Diamond PhD student, Ryan Lithgo, who was based in the MPL, has led to a better understanding of bacterial resistance to silver [2]. The research used a combination of X-ray crystallography (I24), biophysical techniques. including isothermal titration calorimetry and circular dichroism (B23), and quantum mechanics/molecular mechanics (QM/MM) simulations. The research team showed that the metal ion binding site in SilF adapts to fit silver or copper. The implications of this research suggest that Gram-negative bacteria do not appear to have evolved a specific Ag(I) efflux system but instead take advantage of the existing Cu(I) detoxification system. Finally, MPL scientists were part of a team that have developed a versatile approach to high-density microcrystals in lipidic cubic phase for serial crystallography (VIALS) developments in serial crystallography [3] as well as publishing detailed methodology on identifying the most appropriate conditions for membrane protein production using a mammalian expression system [4]. [1] Weckener, M. et al . The lipid linked oligosaccharide polymerase Wzy and its regulating co-polymerase, Wzz, from enterobacterial common antigen biosynthesis form a complex. Open Biol . 13 (3):220373. (2023) DOI: 10.1098/ rsob.220373. [2] Lithgo, R.M. et al . The adaptability of the ion-binding site by the Ag(I)/ Cu(I) periplasmic chaperone SilF. J Biol Chem. 299 (11):105331. (2023) DOI: 10.1016/j.jbc.2023.105331. [3] Gamage, N. et al . High-Throughput Production and Optimization of Membrane Proteins After Expression in Mammalian Cells. Methods Mol Biol . 2652 :79-118. (2023) DOI: 10.1007/978-1-0716-3147-8_5. XChem Capital investment in 2022 and 2023 has enabled the XChem facility to provide even greater support for crystallographic fragment screening and high-throughput structural biology experiments. The increase in throughput of the beamline and an expansion in the sample preparation capabilities allowed collection of more than 45,000 datasets from academic groups and grant-funded research projects in 2023, alongside maintaining a highly active industrial user program. 1,116 X-ray structures from I04-1 have been deposited in the PDB in 2023 and so far in 2024. MPL team circa Aug 2023.