Probing Invisible, Excited Protein States by Non-Uniformly Sampled Pseudo-4D CEST Spectroscopy

被引：24

作者：

Long, Dong ^{[1
,2
]}

Delaglio, Frank ^{[3
]}

Sekhar, Ashok ^{[1
,2
]}

Kay, Lewis E. ^{[1
,2
,4
]}

机构：

[1] Univ Toronto, Dept Mol Genet, Toronto, ON M5S 1A8, Canada

[2] Univ Toronto, Dept Biochem & Chem, Toronto, ON M5S 1A8, Canada

[3] NMR Sci, Campbell, CA 95008 USA

[4] Hosp Sick Children, Program Mol Struct & Funct, Toronto, ON M5G 1X8, Canada

来源：

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | 2015年 / 54卷 / 36期

基金：

加拿大健康研究院;

关键词：

CEST; conformational dynamics; non-uniform sampling; proteins; pseudo-4D NMR spectroscopy; NUCLEAR-MAGNETIC-RESONANCE; ZINC SUPEROXIDE-DISMUTASE; T4; LYSOZYME; RESOLUTION; DYNAMICS; SENSITIVITY; EXCHANGE;

D O I：

10.1002/anie.201504070

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Chemical exchange saturation transfer (CEST) NMR spectroscopy is a powerful tool for studies of slow timescale protein dynamics. Typical experiments are based on recording a large number of 2D data sets and quantifying peak intensities in each of the resulting planes. A weakness of the method is that peaks must be resolved in 2D spectra, limiting applications to relatively small proteins. Resolution is significantly improved in 3D spectra but recording uniformly sampled data is time-prohibitive. Here we describe non-uniformly sampled HNCO-based pseudo-4D CEST that provides excellent resolution in reasonable measurement times. Data analysis is done through fitting in the time domain, without the need of reconstructing the frequency dimensions, exploiting previously measured accurate peak positions in reference spectra. The methodology is demonstrated on several protein systems, including a nascent form of superoxide dismutase that is implicated in neurodegenerative disease.

引用

页码：10507 / 10511

页数：5