title: Exploiting Microbeams for Membrane Protein Structure Determination.
abstract: A reproducible, and sample independent means of predictably obtaining large, well-ordered crystals has proven elusive in macromolecular crystallography. In the structure determination pipeline, crystallisation often proves to be a rate-limiting step, and the process of obtaining even small or badly ordered crystals can prove time-consuming and laborious. This is particularly true in the field of membrane protein crystallography and this is reflected in the limited number of unique membrane protein structures deposited in the protein data bank (less than 650 by June 2016 - http://blanco.biomol.uci.edu/mpstruc ). Over recent years the requirement for, and time and cost associated with obtaining, large crystals has been partially alleviated through the development of beamline instrumentation allowing data collection, and structure solution, from ever-smaller crystals. Advances in several areas have led to a step change in what might be considered achievable during a synchrotron trip over the last decade. This chapter will briefly review the current status of the field, the tools available to ease data collection and processing, and give some examples of exploitation of these for membrane protein microfocus macromolecular crystallography.
authors.affiliation: Diamond Light Source, Harwell Science and Innovation Campus, Didcot, OX11 0DE, UK.
authors.forename: Anna J
authors.initials: AJ
authors.lastname: Warren
authors.affiliation: Diamond Light Source, Harwell Science and Innovation Campus, Didcot, OX11 0DE, UK.
authors.forename: Danny
authors.initials: D
authors.lastname: Axford
authors.affiliation: Diamond Light Source, Harwell Science and Innovation Campus, Didcot, OX11 0DE, UK.
authors.forename: Neil G
authors.initials: NG
authors.lastname: Paterson
authors.affiliation: Diamond Light Source, Harwell Science and Innovation Campus, Didcot, OX11 0DE, UK. robin.owen@diamond.ac.uk.
authors.forename: Robin L
authors.initials: RL
authors.lastname: Owen
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000295:instrumentation / Q000379:methods*; D003625:Data Collection; D030562:Databases, Protein; D019852:Diacylglycerol Kinase / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000737:chemistry; D006801:Humans; D008070:Lipopolysaccharides / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D008857:Microscopy, Interference / Q000379:methods; D008958:Models, Molecular; D018019:Receptors, Corticotropin-Releasing Hormone / Q000737:chemistry; D017356:Synchrotrons / Q000295:instrumentation*; D055114:X-Ray Microtomography / Q000295:instrumentation / Q000379:methods*; D014965:X-Rays; D000099052:CRF Receptor, Type 1
chemical_list: C412459:ABCB10 protein, human; D018528:ATP-Binding Cassette Transporters; D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D008070:Lipopolysaccharides; C475958:LptD protein, E coli; C514179:LptE protein, E coli; D008565:Membrane Proteins; D018019:Receptors, Corticotropin-Releasing Hormone; D000099052:CRF Receptor, Type 1; D019852:Diacylglycerol Kinase
keywords: In situ data collection; Instrumentation; Membrane proteins; Microfocus macromolecular crystallography; X-ray microtomography