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Benchmarking Membrane Protein Detergent Stability for Improving Throughput of High-Resolution X-ray Structures

DOI: 10.1016/j.str.2010.12.001 DOI Help
PMID: 21220112 PMID Help

Authors: Yo Sonoda (Imperial College London) , Simon Newstead (Membrane Protein Laboratory, Imperial College, London) , Nien-jen Hu (Imperial College London; Diamond Light Source) , Yilmaz Alguel (Imperial College MSF; Diamond Light Source; Japan Science and Technology Agency) , Emmanuel Nji (Imperial College London) , Konstantinos Beis (Diamond Light Source) , Shoko Yashiro (Diamond Light Source; Japan Science and Technology Agency) , Chiara Lee (Imperial College London) , James Leung (Imperial College London) , Alexander D. Cameron (Imperial College London) , Bernadette Byrne (Imperial College London) , So Iwata (Diamond Light Source; Japan Science and Technology Agency; Imperial College London) , David Drew (Imperial College London)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Structure , VOL 19 (1) , PAGES 17-25

State: Published (Approved)
Published: January 2011

Open Access Open Access

Abstract: Obtaining well-ordered crystals is a major hurdle to X-ray structure determination of membrane proteins. To facilitate crystal optimization, we investigated the detergent stability of 24 eukaryotic and prokaryotic membrane proteins, predominantly transporters, using a fluorescent-based unfolding assay. We have benchmarked the stability required for crystallization in small micelle detergents, as they are statistically more likely to lead to high-resolution structures. Using this information, we have been able to obtain well-diffracting crystals for a number of sodium and proton-dependent transporters. By including in the analysis seven membrane proteins for which structures are already known, AmtB, GlpG, Mhp1, GlpT, EmrD, NhaA, and LacY, it was further possible to demonstrate an overall trend between protein stability and structural resolution. We suggest that by monitoring membrane protein stability with reference to the benchmarks described here, greater efforts can be placed on constructs and conditions more likely to yield high-resolution structures.

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: Membrane Protein Laboratory
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Added On: 16/03/2011 09:48

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