Assignment of Backbone Resonances for Larger Proteins Using the 13C-1H Coherence of a 1H
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- 作者：Toshio Yamazaki ; Hidehito Tochio ; Junichi Furui ; Saburo Aimoto ; and Yoshimasa Kyogoku
- 刊名：Journal of the American Chemical Society
- 出版年：1997
- 出版时间：February 5, 1997
- 年：1997
- 卷：119
- 期：5
- 页码：872 - 880
- 全文大小：420K
- 年卷期：v.119,no.5(February 5, 1997)
- ISSN：1520-5126
文摘
A suite of triple resonance NMR experiments for theassignment of backbone resonances of a largerprotein using selectively 1H
labeling to asample uniformly labeled with 13C, 15N, and2H is described. The relaxationtime of 1H-13Czero/double quantum coherence was more than 4 times as long as that of13C single quantumcoherence. Three-dimensional HACAN, HACACB, HACACO, andHACA(N)CO experiments were newly designedto utilize selectively labeled 1H nuclei.HACAN provides intraresidue and sequential connectivities throughamide15N spins. HACACO and HACA(N)CO provideintraresidue and sequential connectivities through13CO spins.HACACB provides connectivity to 13C
,giving the type of amino acid. Long-life1H-13Czero/double quantumcoherence provides high sensitivity in these NMR experiments.Except for a few amino acid type-specific problems,all sequential connectivities were obtained for a test sample of a 98amino acid protein at 10 
C, which rotationallydiffuses with a correlation time of 17 ns, corresponding to an over 30kDa protein at 30-40 C. Zero/double quantumbased triple resonance experiments and 1Hselective labeling provide a new approach for NMR studies onlargerproteins.
labeling to asample uniformly labeled with 13C, 15N, and2H is described. The relaxationtime of 1H-13Czero/double quantum coherence was more than 4 times as long as that of13C single quantumcoherence. Three-dimensional HACAN, HACACB, HACACO, andHACA(N)CO experiments were newly designedto utilize selectively labeled 1H nuclei.HACAN provides intraresidue and sequential connectivities throughamide15N spins. HACACO and HACA(N)CO provideintraresidue and sequential connectivities through13CO spins.HACACB provides connectivity to 13C,giving the type of amino acid. Long-life1H-13Czero/double quantumcoherence provides high sensitivity in these NMR experiments.Except for a few amino acid type-specific problems,all sequential connectivities were obtained for a test sample of a 98amino acid protein at 10 C, which rotationallydiffuses with a correlation time of 17 ns, corresponding to an over 30kDa protein at 30-40 C. Zero/double quantumbased triple resonance experiments and 1Hselective labeling provide a new approach for NMR studies onlargerproteins.