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

Christina M. Szabo, Lori K. Sanders, H. Carl Le, Ellen Y.T. Chien, Eric Oldfield

Research output: Contribution to journalArticlepeer-review

Abstract

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. Copyright (C) 2000 Federation of European Biochemical Societies.

Original languageEnglish (US)
Pages (from-to)25-30
Number of pages6
JournalFEBS Letters
Volume482
Issue number1-2
DOIs
StatePublished - Sep 29 2000

Keywords

  • Chemical shift assignment
  • Cytochrome c
  • Nuclear magnetic resonance
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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