A Dipolar Coupling Based Strategy for Simultaneous Resonance Assignment and Structure Determination of Protein Backbones
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- 作者：Fang Tian ; Homayoun Valafar ; and James H. Prestegard
- 刊名：Journal of the American Chemical Society
- 出版年：2001
- 出版时间：November 28, 2001
- 年：2001
- 卷：123
- 期：47
- 页码：11791 - 11796
- 全文大小：148K
- 年卷期：v.123,no.47(November 28, 2001)
- ISSN：1520-5126
文摘
A new approach for simultaneous protein backbone resonance assignment and structure determinationby NMR is introduced. This approach relies on recent advances in high-resolution NMR spectroscopy thatallow observation of anisotropic interactions, such as dipolar couplings, from proteins partially aligned infield ordered media. Residual dipolar couplings are used for both geometric information and a filter in theassembly of residues in a sequential manner. Experimental data were collected in less than one week on asmall redox protein, rubredoxin, that was 15N enriched but not enriched above 1% natural abundance in 13C.Given the acceleration possible with partial 13C enrichment, the protocol described should provide a veryrapid route to protein structure determination. This is critical for the structural genomics initiative where proteinexpression and structural determination in a high-throughput manner will be needed.