Excited States of Nucleic Acids Probed by Proton Relaxation Dispersion NMR Spectroscopy

被引:35
|
作者
Juen, Michael Andreas [1 ,2 ]
Wunderlich, Christoph Hermann [3 ]
Nussbaumer, Felix [1 ,2 ]
Tollinger, Martin [1 ,2 ]
Kontaxis, Georg [4 ]
Konrat, Robert [4 ]
Hansen, D. Flemming [5 ]
Kreutz, Christoph [1 ,2 ]
机构
[1] Univ Innsbruck, Inst Organ Chem, Innrain 80-82, A-6020 Innsbruck, Austria
[2] Univ Innsbruck, CMBI, Innrain 80-82, A-6020 Innsbruck, Austria
[3] Bachem Amer Inc, 3132 Kashiwa St, Torrance, CA 90505 USA
[4] Univ Vienna, Max F Perutz Labs MFPL, Computat Biol & Biomol NMR, Dr Bohr Gasse 9 VBC 5, A-1030 Vienna, Austria
[5] UCL, Div Biosci, Inst Struct & Mol Biol, Darwin Bldg,Room 612,Gower St, London WC1E 6BT, England
基金
英国生物技术与生命科学研究理事会; 奥地利科学基金会;
关键词
DNA; NMR spectroscopy; proton relaxation dispersion; RNA; stable isotope labeling; HIV-1 NUCLEOCAPSID PROTEIN; H-1-NMR CHEMICAL-SHIFTS; RNA DYNAMICS; CTAR DNA; CONFORMATIONAL STATES; H-1; TIME; NUCLEOTIDES; RECOGNITION; CHAPERONE;
D O I
10.1002/anie.201605870
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
In this work an improved stable isotope labeling protocol for nucleic acids is introduced. The novel building blocks eliminate/minimize homonuclear C-13 and H-1 scalar couplings thus allowing proton relaxation dispersion (RD) experiments to report accurately on the chemical exchange of nucleic acids. Using site-specific H-2 and C-13 labeling, spin topologies are introduced into DNA and RNA that make H-1 relaxation dispersion experiments applicable in a straightforward manner. The novel RNA/DNA building blocks were successfully incorporated into two nucleic acids. The A-site RNA was previously shown to undergo a two site exchange process in the micro-to millisecond time regime. Using proton relaxation dispersion experiments the exchange parameters determined earlier could be recapitulated, thus validating the proposed approach. We further investigated the dynamics of the cTAR DNA, a DNA transcript that is involved in the viral replication cycle of HIV-1. Again, an exchange process could be characterized and quantified. This shows the general applicablility of the novel labeling scheme for H-1 RD experiments of nucleic acids.
引用
收藏
页码:12008 / 12012
页数:5
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