Deuterium incorporation into Escherichia-coli proteins

A neutron-scattering study of DNA-dependent RNA polymerase

H. Lederer, R. P. May, Jørgen Kjems, W. Schaefer, H. L. Crespi, H. Heumann

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Neutron small-angle scattering studies of single protein subunits in a protein-DNA complex require the adjustment of the neutron scattering-length densities of protein and DNA, which is attainable by specific deuteration of the protein. The neutron scattering densities of unlabelled DNA and DNA-dependent RNA polymerase of Escherichia coli match when RNA polymerase is isolated from cells grown in a medium containing 46% D2O and unlabelled glucose as carbon source. Their contrasts vanish simultaneously in a dialysis buffer containing 65% D2O. An expression was evaluated which allows the calculation of the degree of deuteration and match point of any E. coli protein from the D2O content of the growth medium, taking the 2H incorporation into RNA polymerase amino acids to be representative for all amino acids in E. coli proteins. The small-angle scattering results, on which the calculation of the degree of deuteration is based, were confirmed by mass spectrometric measurements.
    Original languageEnglish
    JournalEuropean Journal of Biochemistry
    Volume156
    Issue number3
    Pages (from-to)655-660
    ISSN0014-2956
    Publication statusPublished - 1986

    Cite this

    Lederer, H., May, R. P., Kjems, J., Schaefer, W., Crespi, H. L., & Heumann, H. (1986). Deuterium incorporation into Escherichia-coli proteins: A neutron-scattering study of DNA-dependent RNA polymerase. European Journal of Biochemistry, 156(3), 655-660.
    Lederer, H. ; May, R. P. ; Kjems, Jørgen ; Schaefer, W. ; Crespi, H. L. ; Heumann, H. / Deuterium incorporation into Escherichia-coli proteins : A neutron-scattering study of DNA-dependent RNA polymerase. In: European Journal of Biochemistry. 1986 ; Vol. 156, No. 3. pp. 655-660.
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    title = "Deuterium incorporation into Escherichia-coli proteins: A neutron-scattering study of DNA-dependent RNA polymerase",
    abstract = "Neutron small-angle scattering studies of single protein subunits in a protein-DNA complex require the adjustment of the neutron scattering-length densities of protein and DNA, which is attainable by specific deuteration of the protein. The neutron scattering densities of unlabelled DNA and DNA-dependent RNA polymerase of Escherichia coli match when RNA polymerase is isolated from cells grown in a medium containing 46{\%} D2O and unlabelled glucose as carbon source. Their contrasts vanish simultaneously in a dialysis buffer containing 65{\%} D2O. An expression was evaluated which allows the calculation of the degree of deuteration and match point of any E. coli protein from the D2O content of the growth medium, taking the 2H incorporation into RNA polymerase amino acids to be representative for all amino acids in E. coli proteins. The small-angle scattering results, on which the calculation of the degree of deuteration is based, were confirmed by mass spectrometric measurements.",
    author = "H. Lederer and May, {R. P.} and J{\o}rgen Kjems and W. Schaefer and Crespi, {H. L.} and H. Heumann",
    year = "1986",
    language = "English",
    volume = "156",
    pages = "655--660",
    journal = "European Journal of Biochemistry",
    issn = "0014-2956",
    publisher = "Springer Verlag",
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    Lederer, H, May, RP, Kjems, J, Schaefer, W, Crespi, HL & Heumann, H 1986, 'Deuterium incorporation into Escherichia-coli proteins: A neutron-scattering study of DNA-dependent RNA polymerase', European Journal of Biochemistry, vol. 156, no. 3, pp. 655-660.

    Deuterium incorporation into Escherichia-coli proteins : A neutron-scattering study of DNA-dependent RNA polymerase. / Lederer, H.; May, R. P.; Kjems, Jørgen; Schaefer, W.; Crespi, H. L.; Heumann, H.

    In: European Journal of Biochemistry, Vol. 156, No. 3, 1986, p. 655-660.

    Research output: Contribution to journalJournal articleResearchpeer-review

    TY - JOUR

    T1 - Deuterium incorporation into Escherichia-coli proteins

    T2 - A neutron-scattering study of DNA-dependent RNA polymerase

    AU - Lederer, H.

    AU - May, R. P.

    AU - Kjems, Jørgen

    AU - Schaefer, W.

    AU - Crespi, H. L.

    AU - Heumann, H.

    PY - 1986

    Y1 - 1986

    N2 - Neutron small-angle scattering studies of single protein subunits in a protein-DNA complex require the adjustment of the neutron scattering-length densities of protein and DNA, which is attainable by specific deuteration of the protein. The neutron scattering densities of unlabelled DNA and DNA-dependent RNA polymerase of Escherichia coli match when RNA polymerase is isolated from cells grown in a medium containing 46% D2O and unlabelled glucose as carbon source. Their contrasts vanish simultaneously in a dialysis buffer containing 65% D2O. An expression was evaluated which allows the calculation of the degree of deuteration and match point of any E. coli protein from the D2O content of the growth medium, taking the 2H incorporation into RNA polymerase amino acids to be representative for all amino acids in E. coli proteins. The small-angle scattering results, on which the calculation of the degree of deuteration is based, were confirmed by mass spectrometric measurements.

    AB - Neutron small-angle scattering studies of single protein subunits in a protein-DNA complex require the adjustment of the neutron scattering-length densities of protein and DNA, which is attainable by specific deuteration of the protein. The neutron scattering densities of unlabelled DNA and DNA-dependent RNA polymerase of Escherichia coli match when RNA polymerase is isolated from cells grown in a medium containing 46% D2O and unlabelled glucose as carbon source. Their contrasts vanish simultaneously in a dialysis buffer containing 65% D2O. An expression was evaluated which allows the calculation of the degree of deuteration and match point of any E. coli protein from the D2O content of the growth medium, taking the 2H incorporation into RNA polymerase amino acids to be representative for all amino acids in E. coli proteins. The small-angle scattering results, on which the calculation of the degree of deuteration is based, were confirmed by mass spectrometric measurements.

    M3 - Journal article

    VL - 156

    SP - 655

    EP - 660

    JO - European Journal of Biochemistry

    JF - European Journal of Biochemistry

    SN - 0014-2956

    IS - 3

    ER -