用于定量大型蛋白质中快速交换过程的非共振TROSY（R1 rho - R1）。

Off-resonance TROSY (R1 rho - R1) for quantitation of fast exchange processes in large proteins.

作者信息

Kempf James G, Jung Ju-yeon, Sampson Nicole S, Loria J Patrick

机构信息

Department of Chemistry, Yale University, P.O. Box 208107, New Haven, Connecticut 06520, USA.

出版信息

J Am Chem Soc. 2003 Oct 8;125(40):12064-5. doi: 10.1021/ja037101s.

DOI:10.1021/ja037101s

PMID:14518971

Abstract

Current solution NMR experiments for characterizing conformational exchange processes in large proteins are limited to exchange rates ca. 500-3000 s-1. A TROSY-based constant relaxation time (R1rho - R1) experiment is designed to extend this capability to measure motion with rates up to 105 s-1 in large macromolecules. The experiment combines off-resonance spin-lock rf fields, which provide access to the faster time-scale dynamics, with TROSY coherence selection, which extends the molecular-weight range available for study. When implemented on the 53-kDa dimeric enzyme triosephosphate isomerase, the experiment yielded substantial gains in signal-to-noise (up to 60%) over current experiments at modest static magnetic fields (14.1 T). The TROSY (R1rho - R1) experiment should therefore be of general utility for investigation of fast conformational exchange events in large proteins.

摘要

目前用于表征大蛋白质中构象交换过程的溶液核磁共振实验仅限于大约500 - 3000 s-1的交换速率。一种基于TROSY的恒定弛豫时间（R1rho - R1）实验被设计用于扩展这种能力，以测量大分子中速率高达105 s-1的运动。该实验将提供对更快时间尺度动力学的非共振自旋锁定射频场与扩展可研究分子量范围的TROSY相干选择相结合。当在53 kDa的二聚体酶磷酸丙糖异构酶上实施时，该实验在适度的静磁场（14.1 T）下比当前实验在信噪比方面有显著提高（高达60%）。因此，TROSY（R1rho - R1）实验对于研究大蛋白质中的快速构象交换事件应具有普遍用途。