Observation of Through-Hydrogen-Bond 2hJHC′ in a Perdeuterated Protein

Institute of Structural Biology, Forschungszentrum Jülich, Jülich, 52425, Germany.
Journal of Magnetic Resonance (Impact Factor: 2.51). 11/1999; 140(2):510-2. DOI: 10.1006/jmre.1999.1899
Source: PubMed


It is demonstrated that J connectivity between amide protons and hydrogen-bond-accepting carbonyl carbons can be observed in perdeuterated human ubiquitin. A selective pulse scheme is used to detect these small 2hJHC' interactions in the presence of the much larger through-covalent-bond 2JHC' and 3JHC' couplings. The ratio of the observed through-H-bond correlation intensity and the 2JHC' connectivity observed in a reference spectrum indicates 2hJHC' values of ca. 0.4-0.6 Hz, which are only slightly smaller than the corresponding 3hJNC' values. However, for technical reasons, 2hJHC' couplings are more difficult to measure than 3hJNC'.

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    • "NMR chemical shifts (Marin et al., 2004; Wishart and Sykes, 1994; Wishart et al., 1991, 1992) or automated assignment (Bailey-Kellogg et al., 2000a) can also be used. Hydrogen bonds can be determined by NMR from experimentally recorded data (Cordier et al., 1999; Wang et al., 1999), or, e.g., by using backbone resonance assignment programs such as Jigsaw (Bailey-Kellogg et al., 2000a). The user of our algorithm has a choice, to record either (a) one type of backbone RDC (such as NH RDCs) in two aligning media, or (b) two types of backbone RDCs (such as NH and CH RDCs) in a single medium. "
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