CPMG Relaxation Dispersion
CPMG Relaxation Dispersion
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DOI：
10.1007/978-1-62703-658-0_2
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发表时间：
2014-01-01
期刊：
影响因子：
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通讯作者：
Ishima, Rieko
中科院分区：
文献类型：
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作者：
Ishima, Rieko
核磁共振弛豫对蛋白质的分子运动和内部运动非常敏感。通常通过N-15纵向驰豫速率(R-1)、横向驰豫速率(R-2)和{H-1}-N-15核Overhauser效应(NOE)实验来全面阐明蛋白质动力学，主要是在亚纳秒的时间尺度上。相反，恒定松弛时间R-2弥散实验被应用于表征在毫秒时间尺度上相互转换的蛋白质平衡构象。有关蛋白质局部构象平衡的信息提供了关于蛋白质能量格局的重要见解，也有助于解释分子识别机制。在此，我们描述了一种在溶液中进行N-15Carr-Purcell-Meiom-Gill(CPMG)R-2弥散测量的方案，包括蛋白质制备、逐步实验参数设置和数据分析的第一步。
NMR relaxation is sensitive to molecular and internal motion of proteins. N-15 longitudinal relaxation rate (R-1), transverse relaxation rate (R-2), and {H-1}-N-15 Nuclear Overhauser Effect (NOE) experiments are often performed to globally elucidate protein dynamics, primarily on the sub-nanosecond timescale. In contrast, constant relaxation time R-2 dispersion experiments are applied to characterize protein equilibrium conformations that interconvert on the millisecond timescale. Information on local conformational equilibria of proteins provides important insights about protein energy landscapes and is useful to interpret molecular recognition mechanisms as well. Here, we describe a protocol for performing N-15 Carr-Purcell-Meiboom-Gill (CPMG) R-2 dispersion measurements in solution, including protein preparation, step-by-step experimental parameter settings, and the first step of data analysis.