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Facilitated assignment of adenine H2 resonances in oligonucleotides using homonuclear long-range couplings.

ABSTRACT: The small homonuclear coupling constant between H2 and H8 protons of adenine (0.5 +/- 0.2 Hz) was determined and exploited to assign the H2 proton of adenine residues in DNA oligonucleotides. The utility of this approach is demonstrated for two different DNA sequences in D(2)O, for which we show a rapid and unambiguous assignment of the A H2 resonances.

SUBMITTER: Zhang J

PROVIDER: S-EPMC2889177 | biostudies-literature | 2009 Apr

REPOSITORIES: biostudies-literature

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Facilitated assignment of adenine H2 resonances in oligonucleotides using homonuclear long-range couplings.

Zhang Jin J Spring Alexander A Germann Markus W MW

Journal of the American Chemical Society 20090401 15

The small homonuclear coupling constant between H2 and H8 protons of adenine (0.5 +/- 0.2 Hz) was determined and exploited to assign the H2 proton of adenine residues in DNA oligonucleotides. The utility of this approach is demonstrated for two different DNA sequences in D(2)O, for which we show a rapid and unambiguous assignment of the A H2 resonances. ...[more]

PMID: 19323530

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Project description:Fast magic angle spinning (MAS) NMR spectroscopy is emerging as an essential analytical and structural biology technique. Large resolution and sensitivity enhancements observed under fast MAS conditions enable structural and dynamics analysis of challenging systems, such as large macromolecular assemblies and isotopically dilute samples, using only a fraction of material required for conventional experiments. Homonuclear dipolar-based correlation spectroscopy constitutes a centerpiece in the MAS NMR methodological toolbox, and is used essentially in every biological and organic system for deriving resonance assignments and distance restraints information necessary for structural analysis. Under fast MAS conditions (rotation frequencies above 35-40 kHz), dipolar-based techniques that yield multi-bond correlations and non-trivial distance information are ineffective and suffer from low polarization transfer efficiency. To overcome this limitation, we have developed a family of experiments, CORD-RFDR. These experiments exploit the advantages of both zero-quantum RFDR and spin-diffusion based CORD methods, and exhibit highly efficient and broadband dipolar recoupling across the entire spectrum, for both short-range and long-range correlations. We have verified the performance of the CORD-RFDR sequences experimentally on a U-(13)C,(15)N-MLF tripeptide and by numerical simulations. We demonstrate applications of 2D CORD-RFDR correlation spectroscopy in dynein light chain LC8 and HIV-1 CA tubular assemblies. In the CORD-RFDR spectra of LC8 acquired at the MAS frequency of 40 kHz, many new intra- and inter-residue correlations are detected, which were not observed with conventional dipolar recoupling sequences. At a moderate MAS frequency of 14 kHz, the CORD-RFDR experiment exhibits excellent performance as well, as demonstrated in the HIV-1 CA tubular assemblies. Taken together, the results indicate that CORD-RFDR experiment is beneficial in a broad range of conditions, including both high and moderate MAS frequencies and magnetic fields.

Dataset Information

Facilitated assignment of adenine H2 resonances in oligonucleotides using homonuclear long-range couplings.

Publications

Facilitated assignment of adenine H2 resonances in oligonucleotides using homonuclear long-range couplings.

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