Palmer Arthur G, Grey Michael J, Wang Chunyu
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York 10032, USA.
Methods Enzymol. 2005;394:430-65. doi: 10.1016/S0076-6879(05)94018-4.
Transverse relaxation optimized NMR spectroscopy (TROSY) techniques for (1)H-(15)N backbone amide moieties and for (13)CH(3) methyl groups have permitted the development of Hahn spin echo and Carr-Purcell-Meiboom-Gill (CPMG) experiments for characterizing chemical exchange kinetic phenomena on microsecond-millisecond time scales in proteins with molecular masses >50 kDa. This chapter surveys the theoretical bases for TROSY in spin systems subject to chemical exchange linebroadening, the experimental methods that have been developed to quantitatively characterize chemical exchange in large proteins, and the emerging applications to triose phosphate isomerase, hemoglobin, and malate synthase G, with molecular masses ranging from 54 to 82 kDa.
用于（1）H-（15）N主链酰胺基团和（13）CH₃甲基基团的横向弛豫优化核磁共振光谱（TROSY）技术，使得针对分子量>50 kDa的蛋白质中微秒至毫秒时间尺度上的化学交换动力学现象进行表征的哈恩自旋回波和卡尔-珀塞尔-梅博姆-吉尔（CPMG）实验得以发展。本章概述了自旋系统中受化学交换线展宽影响的TROSY的理论基础、已开发用于定量表征大蛋白质中化学交换的实验方法，以及在分子量范围为54至82 kDa的磷酸丙糖异构酶、血红蛋白和苹果酸合酶G上的新兴应用。
