Probing slowly exchanging protein systems via 13Cα-CEST: monitoring folding of the Im7 protein

被引：21

作者：

Hansen, Alexandar L. ^{[1
,2
,3
]}

Bouvignies, Guillaume ^{[1
,2
,3
]}

Kay, Lewis E. ^{[1
,2
,3
,4
]}

机构：

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

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

[3] Univ Toronto, Dept Chem, Toronto, ON M5S 1A8, Canada

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

来源：

JOURNAL OF BIOMOLECULAR NMR | 2013年 / 55卷 / 03期

基金：

加拿大自然科学与工程研究理事会; 美国国家科学基金会; 加拿大健康研究院;

关键词：

Protein dynamics; Slow chemical exchange; Chemical exchange saturation transfer (CEST); Protein folding; Im7; DISPERSION NMR-SPECTROSCOPY; TIME-SCALE DYNAMICS; CHEMICAL-SHIFTS; C-ALPHA; ACCURATE MEASUREMENT; MOLECULAR-DYNAMICS; PULSE SEQUENCES; RELAXATION; STATES; INTERMEDIATE;

D O I：

10.1007/s10858-013-9711-4

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A C-13(alpha) chemical exchange saturation transfer based experiment is presented for the study of protein systems undergoing slow interconversion between an 'observable' ground state and one or more 'invisible' excited states. Here a labeling strategy whereby [2-C-13]-glucose is the sole carbon source is exploited, producing proteins with C-13 at the C-alpha position, while the majority of residues remain unlabeled at CO or C-beta. The new experiment is demonstrated with an application to the folding reaction of the Im7 protein that involves an on-pathway excited state. The obtained excited state C-13(alpha) chemical shifts are cross validated by comparison to values extracted from analysis of CPMG relaxation dispersion profiles, establishing the utility of the methodology.

引用

页码：279 / 289

页数：11

共 76 条

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