Benchmarking membrane protein detergent stability for improving throughput of high-resolution X-ray structures.
Benchmarking membrane protein detergent stability for improving throughput of high-resolution X-ray structures.
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DOI：
10.1016/j.str.2010.12.001
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发表时间：
2011-01-12
期刊：
影响因子：
5.7
通讯作者：
Drew, David
中科院分区：
文献类型：
--
作者：
Sonoda, Yo;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
获得有序晶体是膜蛋白X射线结构测定的主要障碍。为了便于晶体优化，我们研究了洗涤剂的稳定性24真核和原核细胞膜蛋白，主要是转运蛋白，使用基于荧光的展开分析。我们已经对小胶束洗涤剂中结晶所需的稳定性进行了基准测试，因为它们在统计学上更有可能导致高分辨率结构。利用这些信息，我们已经能够获得一些钠和质子依赖性转运蛋白的衍射晶体。通过在分析中包括7种结构已知的膜蛋白，AmtB、GlpG、Mhp1、GlpT、EmrD、NhaA和LacY，可以进一步证明蛋白质稳定性和结构分辨率之间的总体趋势。我们建议，通过监测膜蛋白的稳定性与这里描述的基准，更大的努力可以放在更有可能产生高分辨率的结构的结构和条件。膜蛋白稳定性是蛋白质固有的，而不是去污剂特异性的膜蛋白在LDAO中稳定更可能产生衍射良好的晶体真核细胞膜蛋白在12 M中的稳定性比原核细胞蛋白低3倍
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. ► Benchmarked the stability required for crystallization in small sized detergents ► Membrane protein stability is inherent to the protein rather than detergent specific ► Membrane proteins stable in LDAO are more likely to yield well-diffracting crystals ► Eukaryotic membrane proteins are 3-fold less stable in 12M than prokaryotic proteins
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影响因子：
14.8
作者：
Drew, David;Newstead, Simon;Iwata, So
通讯作者：
Iwata, So
影响因子：
56.9
作者：
Daley, DO;Rapp, M;von Heijne, G
通讯作者：
von Heijne, G
影响因子：
56.9
作者：
Dutzler, R;Campbell, EB;MacKinnon, R
通讯作者：
MacKinnon, R
DOI：
10.1126/science.1168750
发表时间：
2009-03-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Aller SG;Yu J;Ward A;Weng Y;Chittaboina S;Zhuo R;Harrell PM;Trinh YT;Zhang Q;Urbatsch IL;Chang G
通讯作者：
Chang G
影响因子：
56.9
作者：
Long, SB;Campbell, EB;MacKinnon, R
通讯作者：
MacKinnon, R