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Sequential resonance assignment from two-dimensional inter- and intra-residue 15N–1H correlation spectra

✍ Scribed by Permi Perttu; Annila Arto


Publisher
John Wiley and Sons
Year
2001
Tongue
English
Weight
392 KB
Volume
39
Category
Article
ISSN
0749-1581

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✦ Synopsis


Sequential assignment of amide resonances in proteins and peptides can conveniently be derived from twodimensional inter-residue 15 N i -1 H N .i-1/ and intra-residue 15 N i -1 H N i correlation spectra. The inter-residue 15 N i -1 H N .i-1/ correlation spectrum is generated by recording the 15 N i frequency evolution indirectly and subsequently transferring the magnetization to 1 H N .i-1/ of the preceding residue for direct detection. The flow of coherence is established by the (H)N(COCAHA)-TOCSY pulse sequence. Following the path from intra-residue correlation via inter-residue correlation to the next intra-residue correlation results in sequential assignment in two dimensions. This kind of assignment protocol is most amenable to re-establishing sequential assignments of target proteins perturbed by binding of ligands or alternatively signals of peptide ligands can be traced in studies of structure-function relationships by NMR. Copyright