Abstract
Recent studies have shown that lysine side-chain NH3 + groups are excellent probes for NMR investigations of dynamics involving hydrogen bonds and ion pairs relevant to protein function. However, due to rapid hydrogen exchange, observation of 1H-15N NMR cross peaks from lysine NH3 + groups often requires use of a relatively low temperature, which renders difficulty in resonance assignment. Here we present an effective strategy to assign 1H and 15N resonances of NH3 + groups at low temperatures. This strategy involves two new 1H/13C/15N triple-resonance experiments for lysine side chains. Application to a protein-DNA complex is demonstrated.
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Acknowledgments
We thank Tianzhi Wang for maintenance of the spectrometers at UTMB NMR facility and Dan Nguyen for language editing. This work was supported by Grant R01GM105931 from the National Institutes of Health (to J.I.).
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Esadze, A., Zandarashvili, L. & Iwahara, J. Effective strategy to assign 1H-15N heteronuclear correlation NMR signals from lysine side-chain NH3 + groups of proteins at low temperature. J Biomol NMR 60, 23–27 (2014). https://doi.org/10.1007/s10858-014-9854-y
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DOI: https://doi.org/10.1007/s10858-014-9854-y