An intraresidual i(HCA)CO(CA)NH experiment for the assignment of main-chain resonances in 15N, 13C labeled proteins
Authors:
- 19 September 2009
- journal article
- research article
- Published by Springer Nature in Journal of Biomolecular NMR
- Vol. 45 (3) , 301-310
Abstract
An improved pulse sequence, intraresidual i(HCA)CO(CA)NH, is described for establishing solely 13C′(i), 15N(i), 1HN(i) connectivities in uniformly 15N/13C-labeled proteins. In comparison to the “out-and-back” style intra-HN(CA)CO experiment, the new pulse sequence offers at least two-fold higher experimental resolution in the 13C′ dimension and on average 1.6 times higher sensitivity especially for residues in α-helices. Performance of the new experiment was tested on a small globular protein ubiquitin and an intrinsically unfolded 110-residue cancer/testis antigen CT16/PAGE5. Use of intraresidual i(HCA)CO(CA)NH experiment in combination with the established HNCO experiment was crucial for the assignment of highly disordered CT16.Keywords
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