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1H-filtered correlation experiments for assignment and determination of coupling constants in backbone labelled proteins
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
Affiliation:(1) School of Biochemistry and Molecular Biology, University of Leeds, LS2"thinsp"9JT"thinsp", U.K;(2) Martek Biosciences, 6480 Dobbin Road, Columbia, MD, 21045, U.S.A
Abstract:The implementation of [13Cagr,13Cprime,15N,2Hagr] 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 1JCagr,Cbeta couplings enables us to measure 1JCagr,Cprime scalar and 1DCagr,Cprime residual dipolar coupling constants using modified HNCA experiments which do not suffer from sensitivity losses characteristic for 13C constant time experiments.
Keywords:backbone labelling  HNCA  HN(CO)CA  one-bond coupling  ubiquitin
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