用于测量均匀标记寡糖中质子-质子和碳-碳剩余偶极耦合的核磁共振实验。

NMR experiments for the measurement of proton-proton and carbon-carbon residual dipolar couplings in uniformly labelled oligosaccharides.

作者信息

Martin-Pastor Manuel, Canales-Mayordomo Angeles, Jiménez-Barbero Jesús

机构信息

Laboratorio Integral de Estructura de Biomoléculas Jose. R. Carracido, Unidade de Resonancia Magnética, RIAIDT, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, Spain.

出版信息

J Biomol NMR. 2003 Aug;26(4):345-53. doi: 10.1023/a:1024096807537.

DOI:10.1023/a:1024096807537

PMID:12815261

Abstract

A 2D-HSQC-carbon selective/proton selective-constant time COSY, 2D-HSQC-(sel C, sel H)-CT COSY experiment, which is applicable to uniformly (13)C isotopically enriched samples (U-(13)C) of oligosaccharides or oligonucleotides is proposed for the measurement of proton-proton RDC in crowded regions of 2D-spectra. In addition, a heteronuclear constant time-COSY experiment, (13)C-(13)C CT-COSY, is proposed for the measurement of one bond carbon-carbon RDC in these molecules. These two methods provide an extension, to U-(13)C molecules, of the original homonuclear constant time-COSY experiment proposed by Tian et al. (1999) for saccharides. The combination of a number of these RDC with NOE data may provide the method of choice to study oligosaccharide conformation in the free and receptor-bound state.

摘要

提出了一种二维异核单量子相干碳选择性/质子选择性恒时相关谱（2D-HSQC-carbon selective/proton selective-constant time COSY），即二维异核单量子相干（选择性碳，选择性氢）恒时相关谱实验（2D-HSQC-(sel C, sel H)-CT COSY），该实验适用于寡糖或寡核苷酸的均匀（13）C同位素富集样品（U-(13)C），用于测量二维谱拥挤区域的质子-质子剩余偶极耦合（RDC）。此外，还提出了一种异核恒时相关谱实验，即（13）C-（13）C恒时相关谱（(13)C-(13)C CT-COSY），用于测量这些分子中的一键碳-碳RDC。这两种方法将田等人（1999年）为糖类提出的原始同核恒时相关谱实验扩展到了U-(13)C分子。将这些RDC与核Overhauser效应（NOE）数据相结合，可能为研究游离态和受体结合态寡糖构象提供首选方法。