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An efficient NMR approach for obtaining sequence‐specific resonance assignments of larger proteins based on multiple isotopic labeling
Author(s) -
Ikura Mitsuhiko,
Krinks Marie,
Torchia Dennis A.,
Bax Ad
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)81528-v
Subject(s) - chemistry , calmodulin , isotopic labeling , dipeptide , amino acid , protein structure , nuclear magnetic resonance spectroscopy , sequence (biology) , peptide sequence , biochemistry , stereochemistry , enzyme , organic chemistry , gene
By simultaneously incorporating in a protein 13 C‐carbonyl‐ and 15 N‐labeled amino acids with different levels of enrichment, characteristic asymmetric doublet‐like patterns are observed for 15 N nuclei that are directly adjacent to the 13 C 1 ‐labeled residues, providing unambiguous identification of a large number of unique dipeptide fragments of the protein. Additional assignments and qualitative structural information can be obtained from such a selectively labeled protein by recording multiple bond correlation spectra. The procedure is demonstrated for the protein calmodulin, complexed with calcium.