Abstract
Protein–nucleic acid interactions involve intermolecular ion pairs of protein side-chain and DNA or RNA phosphate groups. Using three protein–DNA complexes, we demonstrate that site-specific oxygen-to-sulfur substitution in phosphate groups allows for identification of NMR signals from the protein side-chain NH3 + groups forming the intermolecular ion pairs. A characteristic change in their 1H and 15N resonances upon this modification (i.e., substitution of phosphate to phosphorodithioate) can represent a signature of an intermolecular ion pair. Hydrogen-bond scalar coupling between protein side-chain 15N and DNA phosphorodithiaote 31P nuclei provides direct confirmation of the intermolecular ion pair. The same approach is likely applicable to protein–RNA complexes as well.
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Acknowledgments
This work was supported by Grant R01GM105931 from National Institutes of Health and Grant CHE-1307344 form National Science Foundation. We thank Drs. Xianbin Yang and David Volk for useful discussion.
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Anderson, K.M., Nguyen, D., Esadze, A. et al. A chemical approach for site-specific identification of NMR signals from protein side-chain NH3 + groups forming intermolecular ion pairs in protein–nucleic acid complexes. J Biomol NMR 62, 1–5 (2015). https://doi.org/10.1007/s10858-015-9909-8
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DOI: https://doi.org/10.1007/s10858-015-9909-8