Lanthanide Tagging of Oligonucleotides to Nucleobase for Paramagnetic NMR.

ABSTRACT: Although lanthanide tags, which have large anisotropic magnetic susceptibilities, have already been introduced to enrich NMR parameters by long-range pseudoconact shifts (PCSs) and residual dipolar couplings (RDCs) of proteins, their application to nucleotides has so far been limited to one previous report, due to the high affinities of lanthanides for the phosphodiester backbone of nucleotides and difficult organic synthesis. Herein, we report successful attachment of a lanthanide tag to a chemically synthesized oligonucleotide via a disulfide bond. NMR experiments reveal PCSs of up to 1?ppm and H-H RDCs of up to 8?Hz at 950?MHz. Although weaker magnetic alignment was achieved than with proteins, the paramagnetic data could be fitted to the known structure of the DNA, taking the mobility of the tag into account. While further rigidification of the tag is desirable, this tag could also be used to measure heteronuclear RDCs of ¹³ C,¹⁵ N-labeled chemically synthesized DNA and RNA.

SUBMITTER: Taubert S 

PROVIDER: S-EPMC7754328 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature