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

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

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

Andrew J Dingley,Lydia Nisius,Florence Cordier,Stephan Grzesiek,

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.

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

AJ Dingley,L Nisius,F Cordier,S Grzesiek,

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

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Nature protocols

VOLUME: 3

Page Numbers: 242-8

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ISSN: 1750-2799

DAY: 15

MONTH: 02

YEAR: 2008

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

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LANGUAGE: eng

NlmUniqueID: 101284307

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AFFILIATION: Department of Chemistry and School of Biological Sciences, The University of Auckland, Private Bag 92019, Auckland 1142, New Zealand.

Country: England

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MEDLINETA: Nat Protoc

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