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

doi:10.1016/S0014-5793(00)02032-9

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

Chemistry

Research output: Contribution to journal › Article › peer-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 language	English (US)
Pages (from-to)	25-30
Number of pages	6
Journal	FEBS Letters
Volume	482
Issue number	1-2
DOIs	https://doi.org/10.1016/S0014-5793(00)02032-9
State	Published - 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

Online availability

10.1016/S0014-5793(00)02032-9

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Cite this

@article{39ccf567960046f7b6230e47c75fb09e,

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

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.",

keywords = "Chemical shift assignment, Cytochrome c, Nuclear magnetic resonance, Saccharomyces cerevisiae",

author = "Szabo, \{Christina M.\} and Sanders, \{Lori K.\} and Le, \{H. Carl\} and Chien, \{Ellen Y.T.\} and Eric Oldfield",

note = "We are grateful to Professor A.G. Mauk (University of British Columbia) for providing the pBTR2 plasmid and to Professor G.S. Rule (Carnegie Mellon University) who provided the HBHA(CBCA)NH pulse sequence file. We thank Dr. Feng Lin (Univesity of Illinois at Urbana-Champaign) for assistance with experimental NMR setup. NMR spectra were obtained in the Varian Oxford Instrument Center for Excellence in NMR Laboratory. Funding for this instrumentation was provided in part from the W.M. Keck Foundation, the National Institutes of Health (PHS 1 S10 RR10444-01), and the National Science Foundation (NSF CHE 96-10502). The National Center for Supercomputing Applications computational time was funded in part by the National Computational Science Alliance, NSF Grant MCA-98N039N. This work was supported by the United States Public Health Service (National Institutes of Health Grants HL-19481 and GM-50694).",

year = "2000",

month = sep,

day = "29",

doi = "10.1016/S0014-5793(00)02032-9",

language = "English (US)",

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journal = "FEBS Letters",

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TY - JOUR

T1 - Expression of doubly labeled Saccharomyces cerevisiae iso-1 ferricytochrome c and 1H, 13C and 15N chemical shift assignments by multidimensional NMR

AU - Szabo, Christina M.

AU - Sanders, Lori K.

AU - Le, H. Carl

AU - Chien, Ellen Y.T.

AU - Oldfield, Eric

N1 - We are grateful to Professor A.G. Mauk (University of British Columbia) for providing the pBTR2 plasmid and to Professor G.S. Rule (Carnegie Mellon University) who provided the HBHA(CBCA)NH pulse sequence file. We thank Dr. Feng Lin (Univesity of Illinois at Urbana-Champaign) for assistance with experimental NMR setup. NMR spectra were obtained in the Varian Oxford Instrument Center for Excellence in NMR Laboratory. Funding for this instrumentation was provided in part from the W.M. Keck Foundation, the National Institutes of Health (PHS 1 S10 RR10444-01), and the National Science Foundation (NSF CHE 96-10502). The National Center for Supercomputing Applications computational time was funded in part by the National Computational Science Alliance, NSF Grant MCA-98N039N. This work was supported by the United States Public Health Service (National Institutes of Health Grants HL-19481 and GM-50694).

PY - 2000/9/29

Y1 - 2000/9/29

N2 - 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.

AB - 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.

KW - Chemical shift assignment

KW - Cytochrome c

KW - Nuclear magnetic resonance

KW - Saccharomyces cerevisiae

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U2 - 10.1016/S0014-5793(00)02032-9

DO - 10.1016/S0014-5793(00)02032-9

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