Department of Structural and Computational Biology, Max F. Perutz Laboratories, University of Vienna, Campus Vienna Biocenter 5, A-1030 Vienna, Austria.
J Am Chem Soc. 2010 Feb 10;132(5):1480-1. doi: 10.1021/ja910098s.
A novel NMR method is demonstrated for the investigation of protein ligand interactions. In this approach an adiabatic fast passage pulse, i.e. a long, weak pulse with a linear frequency sweep, is used to probe (1)H-(1)H NOEs. During the adiabatic fast passage the effective rotating-frame NOE is a weighted average of transverse and longitudinal cross-relaxation contributions that can be tuned by pulse power and frequency sweep rate. It is demonstrated that the occurrence of spin diffusion processes leads to sizable deviations from the theoretical relationship between effective relaxation rate and effective tilt angle in the spin lock frame and can be used to probe protein-ligand binding. This methodology comprises high sensitivity and ease of implementation. The feasibility of this technique is demonstrated with two protein complexes, vanillic acid bound to the quail lipocalin Q83 and NAD(+) and AMP binding to alcohol dehydrogenase (ADH).
一种新的 NMR 方法被用于研究蛋白质-配体相互作用。在该方法中，采用绝热快速通道脉冲（adiabatic fast passage pulse），即长而弱的线性频率扫描脉冲，来探测（1）H-（1）H NOEs。在绝热快速通道过程中，有效转动框架 NOE 是横向和纵向交叉弛豫贡献的加权平均值，可通过脉冲功率和频率扫描速率进行调节。结果表明，自旋扩散过程的发生导致有效弛豫率与自旋锁定框架中有效倾斜角之间的理论关系出现较大偏差，并可用于探测蛋白质-配体结合。该方法具有灵敏度高和易于实施的特点。该技术的可行性已通过两个蛋白质复合物得到验证，分别是鹌鹑卵磷蛋白 Q83 结合香草酸和醇脱氢酶（ADH）结合 NAD（+）和 AMP。
