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

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

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

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

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.

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

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

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

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

Journal: Nature protocols

VOLUME: 3

Page Numbers: 235-41

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ISSN: 1750-2799

DAY: 15

MONTH: 02

YEAR: 2008

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

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LANGUAGE: eng

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AFFILIATION: Department of Structural Biology and Chemistry, Institut Pasteur, 28 Rue du Docteur Roux, 75015 Paris, France.

Country: England

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MEDLINETA: Nat Protoc

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