An efficient NMR approach for obtaining sequence‐specific resonance assignments of larger proteins based on multiple isotopic labeling

FEBS Letters

Bax, Ad³  Torchia, Dennis A.¹  Krinks, Marie²  Ikura, Mitsuhiko³ 

[1] Bone Research Branch, NIDR, National Institutes of Health, Bethesda, MD 20892, USA;Laboratory of Biochemistry, NCI, National Institutes of Health, Bethesda, MD 20892, USA;Laboratory of Chemical Physics, NIDDK, National Institutes of Health, Bethesda, MD 20892, USA

关键词: Two-dimensional NMR;    Isotopic labeling;    Sequential assignment;    Calmodulin;    Central helix;   

DOI  :  10.1016/0014-5793(90)81528-V

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

By simultaneously incorporating in a protein ¹³C-carbonyl- and ¹⁵N-labeled amino acids with different levels of enrichment, characteristic asymmetric doublet-like patterns are observed for ¹⁵N nuclei that are directly adjacent to the ¹³C₁-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.

【 授权许可】

Unknown