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PMID: 21220112 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Benchmarking membrane protein detergent stability for improving throughput of high-resolution X-ray structures.

Structure (London, England : 1993) ·Vol. 19 ·No. 1 ·2011-01-12 ·Pages 17-25

Sonoda Y, Newstead S, Hu NJ, Alguel Y, Nji E, Beis K, Yashiro S, Lee C, Leung J, Cameron AD, Byrne B, Iwata S, Drew D

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.

MeSH Terms
Crystallography, X-Ray/methods,standards Detergents/chemistry Green Fluorescent Proteins/chemistry,genetics Membrane Proteins/chemistry,genetics Membrane Transport Proteins/chemistry,genetics Protein Conformation Protein Stability Protein Unfolding Recombinant Fusion Proteins/chemistry,genetics
Chemicals
Detergents Membrane Proteins Membrane Transport Proteins Recombinant Fusion Proteins Green Fluorescent Proteins
Authors & Affiliations
13 authors, click to expand affiliations / ORCID
Sonoda Yo
Division of Molecular Biosciences, Membrane Protein Crystallography Group, Imperial College, London SW7 2AZ, UK.
Newstead Simon
Hu Nien-Jen
Alguel Yilmaz
Nji Emmanuel
Beis Konstantinos
Yashiro Shoko
Lee Chiara
Leung James
Cameron Alexander D
Byrne Bernadette
Iwata So
Drew David
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Article Info
Journal
Structure (London, England : 1993)
Abbr.
Structure
ISSN
1878-4186
DOI
10.1016/j.str.2010.12.001
Published
2011-01-12
Pages
17-25
Language
English
Region
United States
NLM ID
101087697
PMCID
PMC3111809
Subset
IM
Grants
Medical Research Council · G0900399 · United Kingdom
Wellcome Trust · WT089809/Z/09/Z · United Kingdom
Biotechnology and Biological Sciences Research Council · BBS/B/14418 · United Kingdom
Biotechnology and Biological Sciences Research Council · BB/G023425/1 · United Kingdom
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1016/j.str.2010.12.001 PMC Full Text View journal details
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