title: Membrane protein structure determination - the next generation.
abstract: The field of Membrane Protein Structural Biology has grown significantly since its first landmark in 1985 with the first three-dimensional atomic resolution structure of a membrane protein. Nearly twenty-six years later, the crystal structure of the beta2 adrenergic receptor in complex with G protein has contributed to another landmark in the field leading to the 2012 Nobel Prize in Chemistry. At present, more than 350 unique membrane protein structures solved by X-ray crystallography (http://blanco.biomol.uci.edu/mpstruc/exp/list, Stephen White Lab at UC Irvine) are available in the Protein Data Bank. The advent of genomics and proteomics initiatives combined with high-throughput technologies, such as automation, miniaturization, integration and third-generation synchrotrons, has enhanced membrane protein structure determination rate. X-ray crystallography is still the only method capable of providing detailed information on how ligands, cofactors, and ions interact with proteins, and is therefore a powerful tool in biochemistry and drug discovery. Yet the growth of membrane protein crystals suitable for X-ray diffraction studies amazingly remains a fine art and a major bottleneck in the field. It is often necessary to apply as many innovative approaches as possible. In this review we draw attention to the latest methods and strategies for the production of suitable crystals for membrane protein structure determination. In addition we also highlight the impact that third-generation synchrotron radiation has made in the field, summarizing the latest strategies used at synchrotron beamlines for screening and data collection from such demanding crystals. This article is part of a Special Issue entitled: Structural and biophysical characterisation of membrane protein-ligand binding.
authors.affiliation: Department of Life Sciences, Imperial College London, London SW7 2AZ, UK; Membrane Protein Laboratory, Diamond Light Source, Harwell Science and Innovation Campus, Didcot OX11 0DE, UK; Research Complex at Harwell Rutherford Appleton Laboratory, Harwell Science and Innovation Campus, Didcot OX11 0FA, UK. Electronic address: i.moraes@imperial.ac.uk.
authors.forename: Isabel
authors.initials: I
authors.lastname: Moraes
authors.forename: Gwyndaf
authors.initials: G
authors.lastname: Evans
authors.forename: Juan
authors.initials: J
authors.lastname: Sanchez-Weatherby
authors.forename: Simon
authors.initials: S
authors.lastname: Newstead
authors.forename: Patrick D Shaw
authors.initials: PD
authors.lastname: Stewart
mesh_terms: D018360:Crystallography, X-Ray; D003902:Detergents / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D011487:Protein Conformation; D017356:Synchrotrons
chemical_list: D003902:Detergents; D008565:Membrane Proteins
keywords: Crystal dehydration; Crystal seeding; In situ data collection; Macromolecular crystallography; Membrane protein; XFEL