Orekhov Vladislav Yu, Korzhnev Dmitry M, Kay Lewis E
Swedish NMR Center at Göteborg University, Box 465, 40530 Göteborg, Sweden.
J Am Chem Soc. 2004 Feb 18;126(6):1886-91. doi: 10.1021/ja038620y.
TROSY-based NMR relaxation dispersion experiments that measure the decay of double- and zero-quantum (1)H-(15)N coherences as a function of applied (1)H and (15)N radio frequency (rf) fields are presented for studying millisecond dynamic processes in proteins. These experiments are complementary to existing approaches that measure dispersions of single-quantum (15)N and (1)H magnetization. When combined, data from all four coherences provide a more quantitative picture of dynamics, making it possible to distinguish, for example, between two-site and more complex exchange processes. In addition, a TROSY-based pulse scheme is described for measuring the relaxation of amide (1)H single-quantum magnetization, obtained by a simple modification of the multiple-quantum experiments. The new methodology is applied to a point mutant of the Fyn SH3 domain that exchanges between folded and unfolded states at 25 degrees C.
本文介绍了基于TROSY的核磁共振弛豫色散实验，该实验通过测量双量子和零量子（1）H-（15）N相干性随所施加的（1）H和（15）N射频（rf）场的衰减，来研究蛋白质中的毫秒级动力学过程。这些实验是对测量单量子（15）N和（1）H磁化色散的现有方法的补充。将所有四种相干性的数据结合起来，可以提供更定量的动力学图像，例如，能够区分双位点和更复杂的交换过程。此外，还描述了一种基于TROSY的脉冲序列，用于测量酰胺（1）H单量子磁化的弛豫，该序列是通过对多量子实验进行简单修改而获得的。这种新方法应用于Fyn SH3结构域的一个点突变体，该突变体在25℃下在折叠态和未折叠态之间进行交换。
