Assignment of the 15N NMR spectra of reduced and oxidized Escherichia coli thioredoxin

FEBS Letters

Dyson, H.Jane¹  Krause, Günter²  Chandrasekhar, K.¹  Holmgren, Arne² 

[1] Department of Molecular Biology, Research Institute of Scripps Clinic, La Jolla, CA 92037, USA;Department of Physiological Chemistry, Karolinska Institutet, Box 60400, S-10401 Stockholm, Sweden

关键词: Thioredoxin;    Heteronuclear NMR;    Redox protein;    Protein conformation;   

DOI  :  10.1016/0014-5793(91)80679-W

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

As a necessary first step in the use of heteronuclear correlated spectra to obtain high resolution solution structures of the protein, assignment of the ¹⁵N NMR spectra of reduced and oxidized Escherichia coli thioredoxin (M _r 12 000) uniformly labeled with ¹⁵N has been performed. The ¹⁵N chemical shifts of backbone amide nitrogen atoms have been determined for both oxidation states of thioredoxin using ¹⁵N-¹H correlated and two-dimensional heteronuclear single-quantum coherence (HSQC) TOCSY and NOESY spectra. The backbone assignments are complete, except for the proline imide nitrogen resonances and include Gly³³, whose amide proton resonance is difficult to observe in homonuclear ¹H spectra. The differences in the ¹⁵N chemical shift between oxidized and reduced thioredoxin, which occur mainly in the vicinity of the two active site cysteines, including residues distant in the amino acid sequence which form a hydrophobic surface close to the active site, are consistent with the differences observed for proton chemical shifts in earlier work on thioredoxin.

【 授权许可】

Unknown