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Direct detection of N-H[...]N hydrogen bonds in biomolecules by NMR spectroscopy.

Abstract : A nuclear magnetic resonance (NMR) experiment is described for the direct detection of N-H[...]N hydrogen bonds (H-bonds) in 15N isotope-labeled biomolecules. This quantitative HNN-COSY (correlation spectroscopy) experiment detects and quantifies electron-mediated scalar couplings across the H-bond (H-bond scalar couplings), which connect magnetically active (15)N nuclei of the H-bond donor and acceptor. Detectable H-bonds comprise the imino H-bonds in canonical Watson-Crick base pairs, many H-bonds in unusual nucleic acid base pairs and H-bonds between protein backbone or side-chain N-H donor and N acceptor moieties. Unlike other NMR observables, which provide only indirect evidence of the presence of H-bonds, the H-bond scalar couplings identify all partners of the H-bond, the donor, the donor proton and the acceptor in a single experiment. The size of the scalar couplings can be related to H-bond geometries and as a time average to H-bond dynamics. The time required to detect the H-bonds is typically less than 1 d at millimolar concentrations for samples of molecular weight < or = approximately 25 kDa. A C15N/13C-labeled potato spindle tuber viroid T1 RNA domain is used as an example to illustrate this procedure.
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Submitted on : Tuesday, March 10, 2009 - 1:44:46 PM
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Andrew J Dingley, Lydia Nisius, Florence Cordier, Stephan Grzesiek. Direct detection of N-H[...]N hydrogen bonds in biomolecules by NMR spectroscopy.. Nature Protocols, Nature Publishing Group, 2008, 3 (2), pp.242-8. ⟨10.1038/nprot.2007.497⟩. ⟨pasteur-00367112⟩



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