Suppr超能文献

大型寡聚 DNA(“核磁共振窗口”)中部分氘代和完全质子化糖残基中质子的 T2 弛豫率差异提供了互补的结构信息。

The differences in the T2 relaxation rates of the protons in the partially-deuteriated and fully protonated sugar residues in a large oligo-DNA ('NMR-window') gives complementary structural information.

作者信息

Agback P, Maltseva T V, Yamakage S I, Nilson F P, Földesi A, Chattopadhyaya J

机构信息

Department of Bioorganic Chemistry, University of Uppsala, Sweden.

出版信息

Nucleic Acids Res. 1994 Apr 25;22(8):1404-12. doi: 10.1093/nar/22.8.1404.

Abstract

Selective incorporation of the stereospecifically deuteriated sugar moieties (> 97 atom % 2H enhancements at H2', H2'', H3' and H5'/5'' sites, approximately 85 atom % 2H enhancement at H4' and approximately 20 atom % 2H enhancement at H1') in DNA and RNA by the 'NMR-window' approach has been shown to solve the problem of the resonance overlap [refs. 1, 2 & 3]. Such specific deuterium labelling gives much improved resolution and sensitivity of the residual sugar proton (i.e. H1' or H4') vicinal to the deuteriated centers (ref. 3). The T2 relaxation time of the residual protons also increases considerably in the partially-deuteriated (shown by underline) sugar residues in dinucleotides [d(CpG), d(GpC), d(ApT), d(TpA)], trinucleotide r(A2'p5'A2'p5'A) and 20-mer DNA duplex 5'd(C1G2C3-G4C5G6C7G8A9A10T11T12C13G14C15G16C17G18C19G20)(2) 3'. The protons with shorter T2 can be filtered away using a number of different NMR experiments such as ROESY, MINSY or HAL. The NOE intensity of the cross-peaks in these experiments includes only straight pathway from H1' to aromatic proton (i-i and i-i + 1) without any spin-diffusion. The volumes of these NOE cross-peaks could be measured with high accuracy as their intensity is 3 to 4 times larger than the corresponding peaks in the fully protonated residues in the normal NOESY spectra. The structural informations thus obtainable from the residual protons in the partially-deuteriated part of the duplex and the fully protonated part in the 'NMR window' can indeed complement each other.

摘要

通过“NMR窗口”方法在DNA和RNA中选择性掺入立体特异性氘代糖部分(H2'、H2''、H3'和H5'/5''位点的氘增强>97原子%,H4'的氘增强约85原子%,H1'的氘增强约20原子%)已被证明可解决共振重叠问题[参考文献1、2和3]。这种特定的氘标记大大提高了与氘代中心相邻的残留糖质子(即H1'或H4')的分辨率和灵敏度(参考文献3)。在二核苷酸[d(CpG)、d(GpC)、d(ApT)、d(TpA)]、三核苷酸r(A2'p5'A2'p5'A)和20聚体DNA双链体5'd(C1G2C3 - G4C5G6C7G8A9A10T11T12C13G14C15G16C17G18C19G20)(2)3'中,部分氘代(下划线表示)糖残基中残留质子的T2弛豫时间也显著增加。可以使用许多不同的NMR实验,如ROESY、MINSY或HAL,滤去T2较短的质子。这些实验中交叉峰的NOE强度仅包括从H1'到芳香族质子的直接路径(i-i和i-i + 1),没有任何自旋扩散。这些NOE交叉峰的体积可以高精度测量,因为它们的强度比正常NOESY光谱中完全质子化残基中的相应峰大3到4倍。因此,从双链体部分氘代部分的残留质子和“NMR窗口”中完全质子化部分可获得的结构信息确实可以相互补充。

相似文献

引用本文的文献

本文引用的文献

1
Nuclear magnetic resonance relaxation of glycogen H1 in solution.
Biochemistry. 1993 Sep 14;32(36):9417-22. doi: 10.1021/bi00087a021.
3
Deuterium labelling in NMR structural analysis of larger proteins.
Q Rev Biophys. 1990 May;23(2):133-74. doi: 10.1017/s0033583500005527.
6
RNA structure and NMR spectroscopy.RNA结构与核磁共振波谱学。
Q Rev Biophys. 1991 Nov;24(4):479-532. doi: 10.1017/s0033583500003875.
8
The effect of selective deuteration on magnetization transfer in larger proteins.
J Biomol NMR. 1992 Mar;2(2):183-94. doi: 10.1007/BF01875529.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验