利用增溶标签克服溶解度限制：在生物分子 NMR 研究中的成功应用。
Overcoming the solubility limit with solubility-enhancement tags: successful applications in biomolecular NMR studies.
机构信息
Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.
出版信息
J Biomol NMR. 2010 Jan;46(1):23-31. doi: 10.1007/s10858-009-9371-6.
Abstract
Although the rapid progress of NMR technology has significantly expanded the range of NMR-trackable systems, preparation of NMR-suitable samples that are highly soluble and stable remains a bottleneck for studies of many biological systems. The application of solubility-enhancement tags (SETs) has been highly effective in overcoming solubility and sample stability issues and has enabled structural studies of important biological systems previously deemed unapproachable by solution NMR techniques. In this review, we provide a brief survey of the development and successful applications of the SET strategy in biomolecular NMR.We also comment on the criteria for choosing optimal SETs, such as for differently charged target proteins, and recent new developments on NMR-invisible SETs.
摘要
尽管 NMR 技术的快速发展显著扩展了可追踪系统的范围，但制备高溶解性和高稳定性的 NMR 适用样品仍然是许多生物系统研究的瓶颈。溶解度增强标签（SET）的应用在克服溶解性和样品稳定性问题方面非常有效，并使以前认为无法通过溶液 NMR 技术进行研究的重要生物系统的结构研究成为可能。在这篇综述中，我们简要介绍了 SET 策略在生物分子 NMR 中的发展和成功应用。我们还评论了选择最佳 SET 的标准，例如针对不同带电的靶蛋白，以及最近关于 NMR 不可见 SET 的新进展。
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本文引用的文献
1
Prion protein library of recombinant constructs for structural biology.
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2
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J Mol Biol. 2009 Mar 6;386(4):1123-37. doi: 10.1016/j.jmb.2009.01.018.
3
Effects of full-length borealin on the composition and protein-protein interaction activity of a binary chromosomal passenger complex.全长 borealin 对二元染色体乘客复合体的组成和蛋白质 - 蛋白质相互作用活性的影响。
Biochemistry. 2009 Feb 17;48(6):1156-61. doi: 10.1021/bi801298j.
4
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