在均匀富集13C的固体蛋白质中的同核13C J去耦

Homo-nuclear 13C J-decoupling in uniformly 13C-enriched solid proteins.

作者信息

Igumenova Tatyana I, McDermott Ann E

机构信息

Department of Chemistry, Columbia University, 3000 Broadway MC 3113, New York, NY 10027, USA.

出版信息

J Magn Reson. 2005 Jul;175(1):11-20. doi: 10.1016/j.jmr.2005.03.007.

DOI:10.1016/j.jmr.2005.03.007

PMID:15949744

Abstract

Recently, we reported an analysis of carbon lineshapes in high resolution solid-state NMR spectra of uniformly 13C-enriched amino acids. Application of a 13C J-decoupling protocol during the carbon chemical shift evolution period allowed us to separate the contribution of the second-order dipolar shift from that of the 13C-13C J-coupling interactions to carbon linewidths. In this work, we have extended this approach to microcrystalline proteins. We describe the performance of the J-decoupling sequence applied to remove homo-nuclear 13C J-couplings in the 13C spectra of ubiquitin. Analysis of the J-decoupling efficiency for C(alpha) and carbonyl protein sites showed that a significant gain in resolution can be achieved.

摘要

最近，我们报道了对均匀富集13C的氨基酸的高分辨率固态NMR谱中碳谱线形状的分析。在碳化学位移演化期间应用13C J-去耦协议，使我们能够将二阶偶极位移的贡献与13C-13C J-耦合相互作用对碳线宽的贡献区分开来。在这项工作中，我们将此方法扩展到微晶蛋白质。我们描述了应用于去除泛素13C谱中同核13C J-耦合的J-去耦序列的性能。对C(α)和羰基蛋白质位点的J-去耦效率分析表明，可以实现分辨率的显著提高。