Antifreeze activity enhancement by site directed mutagenesis on an antifreeze protein from the beetle Rhagium mordax

Dennis Steven Friis, Erlend Kristiansen, Hans Ramløv, Nicolas von Solms

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    Resumé

    The ice binding motifs of insect antifreeze proteins (AFPs) mainly consist of repetitive TxT motifs aligned on a flat face of the protein. However, these motifs often contain non-threonines that disrupt the TxT pattern. We substituted two such disruptive amino acids located in the ice binding face of an AFP from Rhagium mordax with threonine. Furthermore, a mutant with an extra ice facing TxT motif was constructed. These mutants showed enhanced antifreeze activity compared to the wild type at low concentrations. However, extrapolating the data indicates that the wild type will become the most active at concentrations above 270 lmol.
    OriginalsprogEngelsk
    TidsskriftF E B S Letters
    Vol/bind588
    Udgave nummer9
    Sider (fra-til)1767-1772
    ISSN0014-5793
    DOI
    StatusUdgivet - 2 maj 2014

    Emneord

    • Antifreeze protein
    • Site directed mutagenesis
    • Thermal hysteresis

    Citer dette

    Friis, Dennis Steven ; Kristiansen, Erlend ; Ramløv, Hans ; von Solms, Nicolas. / Antifreeze activity enhancement by site directed mutagenesis on an antifreeze protein from the beetle Rhagium mordax. I: F E B S Letters. 2014 ; Bind 588, Nr. 9. s. 1767-1772.
    @article{186a7beb10604cc291276cec52f120f8,
    title = "Antifreeze activity enhancement by site directed mutagenesis on an antifreeze protein from the beetle Rhagium mordax",
    abstract = "The ice binding motifs of insect antifreeze proteins (AFPs) mainly consist of repetitive TxT motifs aligned on a flat face of the protein. However, these motifs often contain non-threonines that disrupt the TxT pattern. We substituted two such disruptive amino acids located in the ice binding face of an AFP from Rhagium mordax with threonine. Furthermore, a mutant with an extra ice facing TxT motif was constructed. These mutants showed enhanced antifreeze activity compared to the wild type at low concentrations. However, extrapolating the data indicates that the wild type will become the most active at concentrations above 270 μmol.",
    keywords = "Antifreeze protein, Site directed mutagenesis, Thermal hysteresis, Antifreeze protein, Site directed mutagenesis, Thermal hysteresis",
    author = "Friis, {Dennis Steven} and Erlend Kristiansen and Hans Raml{\o}v and {von Solms}, Nicolas",
    year = "2014",
    month = "5",
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    doi = "10.1016/j.febslet.2014.03.032",
    language = "English",
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    pages = "1767--1772",
    journal = "F E B S Letters",
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    Antifreeze activity enhancement by site directed mutagenesis on an antifreeze protein from the beetle Rhagium mordax. / Friis, Dennis Steven; Kristiansen, Erlend; Ramløv, Hans; von Solms, Nicolas.

    I: F E B S Letters, Bind 588, Nr. 9, 02.05.2014, s. 1767-1772.

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    TY - JOUR

    T1 - Antifreeze activity enhancement by site directed mutagenesis on an antifreeze protein from the beetle Rhagium mordax

    AU - Friis, Dennis Steven

    AU - Kristiansen, Erlend

    AU - Ramløv, Hans

    AU - von Solms, Nicolas

    PY - 2014/5/2

    Y1 - 2014/5/2

    N2 - The ice binding motifs of insect antifreeze proteins (AFPs) mainly consist of repetitive TxT motifs aligned on a flat face of the protein. However, these motifs often contain non-threonines that disrupt the TxT pattern. We substituted two such disruptive amino acids located in the ice binding face of an AFP from Rhagium mordax with threonine. Furthermore, a mutant with an extra ice facing TxT motif was constructed. These mutants showed enhanced antifreeze activity compared to the wild type at low concentrations. However, extrapolating the data indicates that the wild type will become the most active at concentrations above 270 μmol.

    AB - The ice binding motifs of insect antifreeze proteins (AFPs) mainly consist of repetitive TxT motifs aligned on a flat face of the protein. However, these motifs often contain non-threonines that disrupt the TxT pattern. We substituted two such disruptive amino acids located in the ice binding face of an AFP from Rhagium mordax with threonine. Furthermore, a mutant with an extra ice facing TxT motif was constructed. These mutants showed enhanced antifreeze activity compared to the wild type at low concentrations. However, extrapolating the data indicates that the wild type will become the most active at concentrations above 270 μmol.

    KW - Antifreeze protein

    KW - Site directed mutagenesis

    KW - Thermal hysteresis

    KW - Antifreeze protein

    KW - Site directed mutagenesis

    KW - Thermal hysteresis

    U2 - 10.1016/j.febslet.2014.03.032

    DO - 10.1016/j.febslet.2014.03.032

    M3 - Journal article

    VL - 588

    SP - 1767

    EP - 1772

    JO - F E B S Letters

    JF - F E B S Letters

    SN - 0014-5793

    IS - 9

    ER -