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  • 1
    ISSN: 1573-5001
    Keywords: FKBP12 ; NMR detection ; sensitivity enhancement ; side chain–main chain hydrogen bonds
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We describe the direct observation of very weak side chain–main chain hydrogen bonding interactions in medium-size 13C/15N-labeled proteins with sensitivity-enhanced NMR spectroscopy. Specifically, the remote correlation between the hydrogen acceptor side chain carboxylate carbon 13CO2 δ of glutamate 54 and the hydrogen donor backbone amide 15N of methionine 49 in a 12 kDa protein, human FKBP12, is detected via the trans-hydrogen bond 3h J NCO2δ coupling by employing a novel sensitivity-enhanced HNCO-type experiment, CPD-HNCO. The 3h J NCO2δ coupling constant appears to be even smaller than the average value of backbone 3h J NC′ couplings, consistent with more extensive local dynamics in protein side chains.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5001
    Keywords: A ⋅ (G ⋅ G ⋅ G ⋅ G) ⋅ A hexad ; G ⋅ C ⋅ G ⋅ C tetrad ; G ⋅ G ⋅ G ⋅ G tetrad ; HNN-COSY ; 2hJNN
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Several structural motifs found in nucleic acids involve N-H ... N hydrogen bonds in which the donor hydrogens are broadened to extinction due to chemical or conformational exchange. In such situations, it is impossible to use the well-established HNN-COSY or soft HNN-COSY experiments, which report the presence of the hydrogen bond directly on the donor proton(s). We present a pulse sequence, H(CN)N(H), for alleviating this problem in hydrogen bonds of the type NdH ... Na-CH, in which the donor Nd nitrogen is correlated with the corresponding non-exchangeable C-H proton associated with the acceptor Na nitrogen. In this way, missing NdH ... Na correlations in an HNN-COSY spectrum may be recovered from CH-Nd correlations in the H(CN)N(H) spectrum. By correlating a different set of nuclei relative to the HNN-COSY class of experiments, the H(CN)N(H) experiment also serves to remove ambiguities associated with degeneracies in HNN-COSY spectra. The technique is demonstrated on d(GGAGGAG)4,a quadruplex containing a novel A ⋅ (G ⋅ G ⋅ G ⋅ G) ⋅ A hexad and on d(GGGCAGGT)4, containing a G ⋅ C ⋅ G ⋅ C tetrad, in which missing NH2 ... N7 correlations are retrieved via H8-(N2,N6) correlations in the H(CN)N(H) spectrum.
    Type of Medium: Electronic Resource
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