title: Direct detection of N-H[...]O=C hydrogen bonds in biomolecules by NMR spectroscopy.
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.
authors.affiliation: Department of Structural Biology and Chemistry, Institut Pasteur, 28 Rue du Docteur Roux, 75015 Paris, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Cordier
authors.forename: Lydia
authors.initials: L
authors.lastname: Nisius
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Dingley
authors.forename: Stephan
authors.initials: S
authors.lastname: Grzesiek
mesh_terms: D002247:Carbon Isotopes; D018946:Chemokine CCL5 / Q000737:chemistry; D006860:Hydrogen Bonding; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D025801:Ubiquitin / Q000737:chemistry
chemical_list: D002247:Carbon Isotopes; D018946:Chemokine CCL5; D009587:Nitrogen Isotopes; D011506:Proteins; D025801:Ubiquitin