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Development and application of aromatic [13C, 1H] SOFAST-HMQC NMR experiment for nucleic acids

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Abstract

Higher sensitivity of NMR spectrometers and novel isotopic labeling schemes have ushered the development of rapid data acquisition methodologies, improving the time resolution with which NMR data can be acquired. For nucleic acids, longitudinal relaxation optimization in conjunction with Ernst angle excitation (SOFAST-HMQC) for imino protons, in addition to rendering rapid pulsing, has been demonstrated to yield significant improvements in sensitivity per unit time. Extending such methodology to other spins offers a viable prospect to measure additional chemical shifts, thereby broadening their utilization for various applications. Here, we introduce the 2D [13C, 1H] aromatic SOFAST-HMQC that results in overall sensitivity gain of 1.4- to 1.7-fold relative to the conventional HMQC and can also be extended to yield long-range heteronuclear chemical shifts such as the adenine imino nitrogens N1, N3, N7 and N9. The applications of these experiments range from monitoring real-time biochemical processes, drug/ligand screening, and to collecting data at very low sample concentration and/or in cases where isotopic enrichment cannot be achieved.

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Acknowledgments

This work was supported by the US National Institute for General Medical Sciences (GM103297), National Institutes of Health (NIAID R01AI066975) and an Agilent Thought Leader Award. We thank Dr. Sven G. Hyberts for his discussions on sparse sampling approaches and reconstructing the reduced 2D NMR dataset. We thank Dr. Vivekanandan Subramanian and the Duke NMR center for maintenance and support of the NMR facilities.

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  1. Department of Biochemistry, Duke University, Durham, NC, 27710, USA

    Bharathwaj Sathyamoorthy, Janghyun Lee, Isaac Kimsey, Laura R. Ganser & Hashim Al-Hashimi

  2. Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA

    Janghyun Lee

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  1. Bharathwaj Sathyamoorthy
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  2. Janghyun Lee
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Correspondence to Hashim Al-Hashimi.

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Sathyamoorthy, B., Lee, J., Kimsey, I. et al. Development and application of aromatic [13C, 1H] SOFAST-HMQC NMR experiment for nucleic acids. J Biomol NMR 60, 77–83 (2014). https://doi.org/10.1007/s10858-014-9856-9

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