Research Complex at Harwell 4 articles published in JoVE Biochemistry Crystallization and In Situ Room Temperature Data Collection Using the Crystallization Facility at Harwell and Beamline VMXi, Diamond Light Source James Sandy1, Halina Mikolajek1,2, Amy J. Thompson1, Juan Sanchez-Weatherby1, Michael A Hough1,2 1Diamond Light Source Ltd, Harwell Science and Innovation Campus, 2Research Complex at Harwell, Harwell Science and Innovation Campus We present a protocol for the crystallization of proteins using the crystallization facility in the Research Complex at Harwell and subsequent in situ X-ray crystallographic data collection from crystals within the plates at Diamond's Versatile Macromolecular Crystallography in situ (VMXi) beamline. We describe sample requirements, crystallization protocols, and data collection guidelines. Chemistry Achieving Efficient Fragment Screening at XChem Facility at Diamond Light Source Alice Douangamath*1,2, Ailsa Powell*1,2, Daren Fearon*1,2, Patrick M. Collins1,2, Romain Talon1,2,3, Tobias Krojer3,4, Rachael Skyner1,2, Jose Brandao-Neto1,2, Louise Dunnett1,2, Alexandre Dias1,2, Anthony Aimon1,2,3, Nicholas M. Pearce1,3, Conor Wild3,5, Tyler Gorrie-Stone1, Frank von Delft1,2,3,4,6 1Diamond Light Source Ltd, Harwell Science and Innovation Campus, 2Research Complex at Harwell, Harwell Science and Innovation Campus, 3Structural Genomics Consortium, University of Oxford, 4Centre for Medicines Discovery, University of Oxford, 5Oxford Protein Informatics Group, Department of Statistics, Oxford University, 6Department of Biochemistry, University of Johannesburg This paper describes the complete XChem process for crystal-based fragment screening, starting from applying for access and all subsequent steps to data dissemination. Biochemistry Biochemical and Structural Characterization of the Carbohydrate Transport Substrate-binding-protein SP0092 Simone Culurgioni1,2, Minzhe Tang1,2, David R. Hall1, Martin A. Walsh1,2 1Diamond Light Source, Harwell Science & Innovation Campus, 2Research Complex at Harwell, Harwell Science & Innovation Campus A streamlined protocol for performing an extensive biochemical and structural characterization of a carbohydrate substrate binding protein from Streptococcus pneumoniae is presented. Biology Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli Louise E. Bird1, Heather Rada1, Anil Verma1, Raphael Gasper2,3, James Birch4, Matthew Jennions4, Jan Lӧwe3, Isabel Moraes4, Raymond J. Owens1 1Oxford Protein Production Facility, Research Complex at Harwell, 2Protein Crystallography Group, Ruhr University, 3MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, 4Membrane Protein Laboratory, Diamond Light Source A streamlined approach to screening for the expression of recombinant membrane proteins in Escherichia coli based on fusion to green fluorescent protein is presented.
