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
收藏
页码：203 / 208
页数：6
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