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Differentiation between unlabeled and very-low-level fully 15N,13C-labeled nucleotides for resonance assignments in nucleic acids

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Abstract

Based on different characteristics between unlabeled and fully 15N,13C-labeled nucleotides, we develop a method for unambiguous resonance assignments in nucleic acids following site-specific fully 15N,13C isotope incorporation at very low levels1. The J-couplings between heteronuclei provide for distinction between the NMR signals of the fully labeled nucleotides and those of the natural abundance nucleotides. The method is demonstrated for DNA oligonucleotides2, in the dimeric G-quadruplex [d(GGGTTCAGG)]2and in the 22-nucleotide human telomeric fragment d[AG3(TTAG3)3]. We expect this approach to be useful for selective monitoring of important functional domains and of their interactions in large nucleic acids.

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Authors and Affiliations

  1. Cellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center, New York, NY, 10021, U.S.A.

    Anh Tuân Phan & Dinshaw J. Patel

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  1. Anh Tuân Phan
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  2. Dinshaw J. Patel
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Correspondence to Anh Tuân Phan.

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Phan, A.T., Patel, D.J. Differentiation between unlabeled and very-low-level fully 15N,13C-labeled nucleotides for resonance assignments in nucleic acids. J Biomol NMR 23, 257–262 (2002). https://doi.org/10.1023/A:1020277223482

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