Ian Mitchelle S. de Vera, Mandy E. Blackburn, Luis Galiano, Gail E. Fanucci
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引用次数: 19
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Abstract
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 herein. Curr. Protoc. Protein Sci . 74:17.17.1-17.17.29. © 2013 by John Wiley & Sons, Inc.
利用定位自旋标记(SDSL)测定可溶性蛋白的脉冲EPR距离
脉冲电子顺磁共振(EPR)在结构生物学领域的复兴主要是由于双电子-电子共振(DEER)技术在距离测量方面的最新进展。本单元以HIV-1蛋白酶为模型，通过位点定向自旋标记(SDSL)对可溶性蛋白中的工程半胱氨酸残基进行基于epr的距离测量。为了阐明蛋白质的构象变化，对实验方案进行了优化，并利用现有的数据分析程序推导了距离分布曲线。本文还概述了伪影抑制的实验考虑、样品制备和误差分析。咕咕叫。Protoc。蛋白质科学，74:17.17.1-17.17.29。©2013 by John Wiley &儿子,Inc。
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