Expression of doubly labeled Saccharomyces cerevisiae iso‐1 ferricytochrome c and 1H, 13C and 15N chemical shift assignments by multidimensional NMR

FEBS Letters

Le, H.Carl¹  Szabo, Christina M¹  Chien, Ellen Y.T¹  Oldfield, Eric¹  Sanders, Lori K¹ 

[1] Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, IL 61801, USA

关键词: Cytochrome c;    Nuclear magnetic resonance;    Chemical shift assignment;    Saccharomyces cerevisiae;   

DOI  :  10.1016/S0014-5793(00)02032-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have expressed [U-¹³C,¹⁵N]-labeled Saccharomyces cerevisiae iso-1 cytochrome c C102T;K72A in Escherichia coli with a yield of 11 mg/l of growth medium. Nuclear magnetic resonance (NMR) studies were conducted on the Fe³⁺ form of the protein. We report herein chemical shift assignments for amide ¹H and ¹⁵N, ¹³C°, ¹³C^α, ¹³C^β, ¹H^α and ¹H^β resonances based upon a series of three-dimensional NMR experiments: HNCA, HN(CO)CA, HNCO, HN(CA)CO, HNCACB, HCA(CO)N, HCCH-TOCSY and HBHA(CBCA)NH. An investigation of the chemical shifts of the threonine residues was also made by using density functional theory in order to help solve discrepancies between ¹⁵N chemical shift assignments reported in this study and those reported previously.

【 授权许可】

Unknown