Improved 3D triple resonance experiments, HNN and HN(C)N, for HN and 15N sequential correlations in (13C, 15N) labeled proteins:: Application to unfolded proteins

被引:173
作者
Panchal, SC [1 ]
Bhavesh, NS [1 ]
Hosur, RV [1 ]
机构
[1] Tata Inst Fundamental Res, Dept Chem Sci, Mumbai 400005, India
关键词
HN(C)N; HNN; sequential N-15 correlations; triple resonance experiments; triplets; unfolded proteins;
D O I
10.1023/A:1011239023422
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
Two triple resonance experiments, HNN and HN(C)N, are presented which correlate H-N and N-15 resonances sequentially along the polypeptide chain of a doubly (C-13, N-15) labeled protein. These incorporate several improvements over the previously published sequences for a similar purpose and have several novel features. The spectral characteristics enable direct identification of certain triplets of residues, which provide many starting points for the sequential assignment procedure. The experiments are sensitive and their utility has been demonstrated with a 22 kDa protein under unfolding conditions where most of the standard triple resonance experiments such as HNCA, CBCANH etc. have limited success because of poor amide, C-alpha and C-beta chemical shift dispersions.
引用
收藏
页码:135 / 147
页数:13
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