Triple resonance-based 13Cα and 13Cβ CEST experiments for studies of ms timescale dynamics in proteins

被引：26

作者：

Long, Dong ^{[1
,2
,3
]}

Sekhar, Ashok ^{[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 | 2014年 / 60卷 / 04期

基金：

加拿大健康研究院;

关键词：

CEST; Conformationally excited protein states; C-13(alpha/beta) chemical shifts; Chemical exchange; Fyn SH3; Protein folding; ISOTOPICALLY-ENRICHED PROTEINS; SIDE-CHAIN RESONANCES; NMR CHEMICAL-SHIFTS; C-13/N-15-ENRICHED PROTEINS; HYDROGEN-EXCHANGE; LARGER PROTEINS; BACKBONE AMIDE; SPECTROSCOPY; CONFORMATION; SENSITIVITY;

D O I：

10.1007/s10858-014-9868-5

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 pair of triple resonance based CEST pulse schemes are presented for measuring C-13(alpha) and C-13(beta) chemical shifts of sparsely populated and transiently formed conformers that are invisible to traditional NMR experiments. CEST profiles containing dips at resonance positions of C-13(alpha) or C-13(beta) spins of major (ground) and minor (excited) conformers are obtained in a pseudo 3rd dimension that is generated by quantifying modulations of cross peaks in N-15, H-1(N) correlation spectra. An application to the folding reaction of a G48A mutant of the Fyn SH3 domain is presented, illustrating and validating the methodology.

引用

页码：203 / 208

页数：6

共 31 条

[11] 3-DIMENSIONAL TRIPLE-RESONANCE NMR-SPECTROSCOPY OF ISOTOPICALLY ENRICHED PROTEINS [J].