用于（3）J（HH）测定和明确 NOE 分配的（5'R）/（5'S）-氘标记 DNA 十聚体中 H5'和 H5″的立体专一性分配。

Stereospecific assignment of H5' and H5″ in a (5'R)-/(5'S)-deuterium- labeled DNA decamer for(3) J (HH) determination and unambiguous NOE assignments.

机构信息

Institute for Protein Research, Osaka University, Suita, Osaka, 565, Japan.

出版信息

J Biomol NMR. 1998 Jan;11(1):103-9. doi: 10.1023/A:1008286925516.

PMID:20859785

Abstract

The stereoselective deuterium labeling at the 5' methylene protons of the ribose ring recently developed by Kawashima et al. [1995, Tetrahedron Lett., 36, 6699-6700] enabled the assignment of pro-R and pro-S protons at the 5' position. The deuterium-labeled nucleotides, [(5'S)-(2)H]- and [(5'R)-(2)H]-diastereomers, in an approximate ratio of 2:1, were incorporated in the decamer 5'-d(GCATTAATGC)-3'. Thus, both pro-R and pro-S methylene proton signals without geminal coupling appeared in the NOESY and DQF-COSY spectra. Complete stereospecific assignments and simplified spin systems enabled the determination of 15 (3)J coupling constants between H4' and H5'/H5", and the unambiguous assignment of 135 NOESY cross peaks originating from H4'/H5'/H5" resonances.

摘要

最近，Kawashima 等人开发了一种在核糖环 5'亚甲基质子上进行立体选择性氘标记的方法[1995，四面体快报，36，6699-6700]，从而能够对 5'位的 pro-R 和 pro-S 质子进行分配。氘标记的核苷酸，[(5'S)-(2)H]-和[(5'R)-(2)H]-非对映异构体，以大约 2:1 的比例，被掺入到十聚体 5'-d(GCATTAATGC)-3'中。因此，没有偕二甲基耦合的两个 pro-R 和 pro-S 亚甲基质子信号都出现在 NOESY 和 DQF-COSY 谱中。完整的立体特异性分配和简化的自旋系统使得能够确定 H4'和 H5'/H5"之间的 15 个(3)J 耦合常数，并明确分配来自 H4'/H5'/H5"共振的 135 个 NOESY 交叉峰。