Pulsed EPR distance measurements in soluble proteins by site-directed spin labeling (SDSL).
Pulsed EPR distance measurements in soluble proteins by site-directed spin labeling (SDSL).
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DOI：
10.1002/0471140864.ps1717s74
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发表时间：
2013-11-05
影响因子：
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通讯作者：
Fanucci, Gail E
中科院分区：
文献类型：
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作者：
de Vera, Ian Mitchelle S;Blackburn, Mandy E;Galiano, Luis;Fanucci, Gail E
脉冲电子顺磁共振(EPR)在结构生物学中的复兴集中在使用双电子-电子共振(DER)技术的距离测量方面的最新改进。本单位重点研究基于EPR的距离测量，通过定点定向自旋标记(SDSL)对可溶性蛋白质中的工程半胱氨酸残基进行测量，并以HIV-1蛋白酶为模型。为了阐明蛋白质的构象变化，对实验方案进行了优化，并使用现有的数据分析程序来推导距离分布曲线。文中还概述了伪影抑制的实验考虑、样品制备和误差分析。
The resurgence of pulsed electron paramagnetic resonance (EPR) in structural biology centers on recent improvements in distance measurements using the double electron-electron resonance (DEER) technique. This unit focuses on EPR-based distance measurements by site-directed spin-labeling (SDSL) of engineered cysteine residues in soluble proteins, with HIV-1 protease used as a model. To elucidate conformational changes in proteins, experimental protocols were optimized and existing data analysis programs were employed to derive distance distribution profiles. Experimental considerations, sample preparation and error analysis for artifact suppression are also outlined here.