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

Cordier, Florence and Nisius, Lydia and Dingley, Andrew J. and Grzesiek, Stephan. (2008) Direct detection of N-H[...]O=C hydrogen bonds in biomolecules by NMR spectroscopy. Nature Protocols, 3 (2). pp. 235-241.

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Official URL: http://edoc.unibas.ch/dok/A5258755

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Abstract

A nuclear magnetic resonance (NMR) experiment is described for the direct detection of N-H[...]O=C hydrogen bonds (H-bonds) in 15N and 13C isotope-labeled biomolecules. This quantitative 'long-range' HNCO-COSY (correlation spectroscopy) experiment detects and quantifies electron-mediated scalar couplings across the H-bond (H-bond scalar couplings), which connect the magnetically active (15)N and (13)C nuclei on both sides of the H-bond. Detectable H-bonds comprise the canonical backbone H-bonds in proteins as well as other H-bonds in proteins and nucleic acids with N-H donors and O=C (carbonylic or carboxylic) acceptors. 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. The time required to detect the N-H[...]O=C H-bonds in small proteins (> or = approximately 10 kDa) is typically on the order of 1 d at millimolar concentrations, whereas H-bond detection for larger proteins (> or = approximately 30 kDa) may be possible within several days depending on concentration, isotope composition, magnetic field strength and molecular weight. The proteins ubiquitin (8.6 kDa), dimeric RANTES (2 x 8.5 kDa) and MAP30 (30 kDa) are used as examples to illustrate this procedure.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology (Grzesiek)
UniBasel Contributors:Grzesiek, Stephan
Item Type:Article, refereed
Article Subtype:Research Article
Bibsysno:Link to catalogue
Publisher:Nature Publishing Group
ISSN:1754-2189
e-ISSN:1750-2799
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:28 Feb 2019 10:52
Deposited On:22 Mar 2012 13:32

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