1H-filtered correlation experiments for assignment and determination of coupling constants in backbone labelled proteins |
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Authors: | Alexander W. Giesen Lynnette C. Bae Christa L. Barrett Jason A. Chyba Michael M. Chaykovsky Minn-Chang Cheng Jenny H. Murray Ed J. Oliver Sarah M. Sullivan Jonathan Miles Brown Steve W. Homans |
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Affiliation: | (1) School of Biochemistry and Molecular Biology, University of Leeds, LS29JT, U.K;(2) Martek Biosciences, 6480 Dobbin Road, Columbia, MD, 21045, U.S.A |
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Abstract: | The implementation of [13C,13C,15N,2H] labelled amino acids into proteins allows the acquisition of high resolution triple resonance experiments. We present for the first time resonance assignments facilitated by this new labelling strategy. The absence of 1JC,C couplings enables us to measure 1JC,C scalar and 1DC,C residual dipolar coupling constants using modified HNCA experiments which do not suffer from sensitivity losses characteristic for 13C constant time experiments. |
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Keywords: | backbone labelling HNCA HN(CO)CA one-bond coupling ubiquitin |
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