Rationalizing α-helical membrane protein crystallization
TLDR
A detailed analysis of the crystallization conditions from 121 α‐helical membrane protein structures deposited in the Protein Data Bank is undertaken so that the success of different parameters can be easily compared for different membrane protein families.Abstract:
X-ray crystallography is currently the most successful method for determining the three-dimensional structure of membrane proteins. Nevertheless, growing the crystals required for this technique presents one of the major bottlenecks in this area of structural biology. This is especially true for the α-helical type membrane proteins that are of particular interest due to their medical relevance. To address this problem we have undertaken a detailed analysis of the crystallization conditions from 121 α-helical membrane protein structures deposited in the Protein Data Bank. This information has been analyzed so that the success of different parameters can be easily compared for different membrane protein families. Concurrent with this analysis, we also present the new sparse matrix crystallization screen MemGold.read more
Citations
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Overcoming the challenges of membrane protein crystallography.
TL;DR: Technological advances are emerging for effective expression, solubilisation, purification and crystallisation of membrane proteins, which will lead to a rapid increase in the rate at which membrane protein structures are solved in the near future.
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Molecular basis of nitrate uptake by the plant nitrate transporter NRT1.1
Joanne L. Parker,Simon Newstead +1 more
TL;DR: Comparison with peptide transporters further reveals how the NRT1/PTR family has evolved to recognize diverse nitrogenous ligands, while maintaining elements of a conserved coupling mechanism within this superfamily of nutrient transporter.
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K2P channel gating mechanisms revealed by structures of TREK-2 and a complex with Prozac
Y.Y. Dong,Ashley C. W. Pike,Alexandra Mackenzie,Alexandra Mackenzie,Conor McClenaghan,Prafulla Aryal,L. Dong,A. Quigley,Mariana Grieben,S. Goubin,Shubhashish M.M. Mukhopadhyay,Gian Filippo Ruda,Gian Filippo Ruda,Michael V. Clausen,Lishuang Cao,Paul Brennan,Paul Brennan,N.A. Burgess-Brown,Mark S.P. Sansom,Stephen J. Tucker,Elisabeth P. Carpenter,Elisabeth P. Carpenter +21 more
TL;DR: Crystal structures of the human TREK-2 channel are presented in two conformations and in complex with norfluoxetine, the active metabolite of fluoxettine (Prozac) and a state-dependent blocker of TREK channels, providing an explanation for TREK channel mechanosensitivity, regulation by diverse stimuli, and possible off-target effects of the serotonin reuptake inhibitor Prozac.
Journal ArticleDOI
The MORPHEUS protein crystallization screen
TL;DR: MORPHEUS is an initial protein crystallization screen with a unique organization which integrates components and ligands selected after analysing all crystal structure data deposited with the Protein Data Bank and local data gathered at the MRC Laboratory of Molecular Biology, Cambridge, England.
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Membrane protein structure determination - the next generation.
Isabel Moraes,Gwyndaf Evans,Juan Sanchez-Weatherby,Simon Newstead,Simon Newstead,Patrick D. Shaw Stewart +5 more
TL;DR: This review draws attention to the latest methods and strategies for the production of suitable crystals for membrane protein structure determination, and highlights the impact that third-generation synchrotron radiation has made in the field.
References
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Sparse matrix sampling: a screening method for crystallization of proteins
Jaru Jancarik,Sung-Hou Kim +1 more
TL;DR: A set of screening conditions for initial experiments in protein crystallization has been developed, tested, and is herein presented as discussed by the authors, which are empirically derived based on known or published crystallization conditions of various proteins in the past, so as to sample as large a range of buffer, pH, additive and precipitant variables as possible, using small amounts of proteins.
Journal ArticleDOI
Crystal structure of the human beta2 adrenergic G-protein-coupled receptor.
Søren G. F. Rasmussen,Hee Jung Choi,Daniel M. Rosenbaum,Tong Sun Kobilka,Foon Sun Thian,Patricia C. Edwards,Manfred Burghammer,Venkata R. P. Ratnala,Ruslan Sanishvili,Robert F. Fischetti,Gebhard F. X. Schertler,William I. Weis,Brian K. Kobilka +12 more
TL;DR: In this paper, the human beta2 adrenoceptor (beta2AR) was crystallized in a lipid environment when bound to an inverse agonist and in complex with a Fab that binds to the third intracellular loop.
Journal ArticleDOI
Molecular basis of proton motive force generation: structure of formate dehydrogenase-N.
TL;DR: The structure of the membrane protein formate dehydrogenase-N (Fdn-N), a major component of Escherichia coli nitrate respiration, has been determined and provides critical insights into the proton motive force generation by redox loop, a common mechanism among a wide range of respiratory enzymes.
Journal ArticleDOI
Three-dimensional crystals of a membrane protein complex: The photosynthetic reaction centre from Rhodopseudomonas viridis
TL;DR: Reaction centres of photosynthesis isolated from the photosynthetic bacterium Rhodopseudomonas viridis by a single step of molecular sieve chromatography were crystallized.
Journal ArticleDOI
High-throughput fluorescent-based optimization of eukaryotic membrane protein overexpression and purification in Saccharomyces cerevisiae
TL;DR: This work presents a cost-effective high-throughput approach for rapidly screening membrane proteins that can be overproduced to levels of >1 mg per liter in Saccharomyces cerevisiae, and illustrates the advantage of such an approach, with the purification of monodisperse human and yeast nucleotide-sugar transporters to unprecedented levels.