用于确定骨架标记蛋白质中耦合常数及进行归属的1H滤波相关实验。

1H-filtered correlation experiments for assignment and determination of coupling constants in backbone labelled proteins.

作者信息

Giesen Alexander W, Bae Lynnette C, Barrett Christa L, Chyba Jason A, Chaykovsky Michael M, Cheng Minn-Chang, Murray Jenny H, Oliver Ed J, Sullivan Sarah M, Brown Jonathan Miles, Homans Steve W

机构信息

School of Biochemistry and Molecular Biology, University of Leeds, UK.

出版信息

J Biomol NMR. 2002 Jan;22(1):21-6. doi: 10.1023/a:1013831417615.

DOI:10.1023/a:1013831417615

PMID:11885977

Abstract

The implementation of [13Calpha,13C',15N,2Halpha] labelled amino acids into proteins allows the acquisition of high resolution triple resonance experiments. We present for the first time resonance assignments facilitated by this new labelling strategy. The absence of 1JCalpha,Cbeta couplings enables us to measure 1JCalpha,C' scalar and 1DCalpha,C' residual dipolar coupling constants using modified HNCA experiments which do not suffer from sensitivity losses characteristic for 13C constant time experiments.

摘要

将[13Cα,13C′,15N,2Hα]标记的氨基酸掺入蛋白质中，使得能够进行高分辨率的三共振实验。我们首次展示了由这种新标记策略促成的共振归属。1JCalpha,Cbeta偶合的缺失使我们能够使用改进的HNCA实验来测量1JCalpha,C′标量和1DCalpha,C′残余偶极偶合常数，这些实验不会受到13C恒时实验特有的灵敏度损失的影响。