Low thermodynamic but high kinetic stability of an antifreeze protein from Rhagium mordax

Dennis Steven Friis, Johannes Lørup Johnsen, Erlend Kristiansen, Peter Westh, Hans Ramløv

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    Resumé

    The equilibrium heat stability and the kinetic heat tolerance of a recombinant antifreeze protein (AFP) from the beetle Rhagium mordax (RmAFP1) are studied through differential scanning calorimetry and circular dichroism spectroscopy. In contrast to other insect AFPs studied with this respect, the RmAFP1 has only one disulfide bridge. The melting temperature, Tm, of the protein is determined to be 28.5°C (pH 7.4), which is much lower than most of those reported for AFPs or globular proteins in general. Despite its low melting temperature, both biophysical and activity measurements show that the protein almost completely refolds into the native state after repeated exposure of 70°C. RmAFP1 thus appears to be kinetically stable even far above its melting temperature. Thermodynamically, the insect AFPs seem to be dividable in three groups, relating to their content of disulfide bridges and widths of the ice binding motifs; high melting temperature AFPs (high disulfide content, TxT motifs), low melting temperature but high refolding capability AFPs (one disulfide bridge, TxTxTxT motifs) and irreversibly unfolded AFPs at low temperatures (no disulfide bridges, TxTxTxTxT motifs). The property of being able to cope with high temperature exposures may appear peculiar for proteins which strictly have their effect at subzero temperatures. Different aspects of this are discussed.
    OriginalsprogEngelsk
    TidsskriftProtein Science
    Vol/bind23
    Udgave nummer6
    Sider (fra-til)760-768
    Antal sider9
    ISSN0961-8368
    DOI
    StatusUdgivet - jun. 2014

    Emneord

    • Antifreeze protein
    • Rhagium mordax
    • Stability
    • thermodynamic stability

    Citer dette

    Friis, Dennis Steven ; Johnsen, Johannes Lørup ; Kristiansen, Erlend ; Westh, Peter ; Ramløv, Hans. / Low thermodynamic but high kinetic stability of an antifreeze protein from Rhagium mordax. I: Protein Science. 2014 ; Bind 23, Nr. 6. s. 760-768.
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    abstract = "The equilibrium heat stability and the kinetic heat tolerance of a recombinant antifreeze protein (AFP) from the beetle Rhagium mordax (RmAFP1) are studied through differential scanning calorimetry and circular dichroism spectroscopy. In contrast to other insect AFPs studied with this respect, the RmAFP1 has only one disulfide bridge. The melting temperature, Tm, of the protein is determined to be 28.5°C (pH 7.4), which is much lower than most of those reported for AFPs or globular proteins in general. Despite its low melting temperature, both biophysical and activity measurements show that the protein almost completely refolds into the native state after repeated exposure of 70°C. RmAFP1 thus appears to be kinetically stable even far above its melting temperature. Thermodynamically, the insect AFPs seem to be dividable in three groups, relating to their content of disulfide bridges and widths of the ice binding motifs; high melting temperature AFPs (high disulfide content, TxT motifs), low melting temperature but high refolding capability AFPs (one disulfide bridge, TxTxTxT motifs) and irreversibly unfolded AFPs at low temperatures (no disulfide bridges, TxTxTxTxT motifs). The property of being able to cope with high temperature exposures may appear peculiar for proteins which strictly have their effect at subzero temperatures. Different aspects of this are discussed.",
    keywords = "Antifreeze protein, Rhagium mordax, Stability, thermodynamic stability",
    author = "Friis, {Dennis Steven} and Johnsen, {Johannes L{\o}rup} and Erlend Kristiansen and Peter Westh and Hans Raml{\o}v",
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    Low thermodynamic but high kinetic stability of an antifreeze protein from Rhagium mordax. / Friis, Dennis Steven; Johnsen, Johannes Lørup; Kristiansen, Erlend; Westh, Peter; Ramløv, Hans.

    I: Protein Science, Bind 23, Nr. 6, 06.2014, s. 760-768.

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    TY - JOUR

    T1 - Low thermodynamic but high kinetic stability of an antifreeze protein from Rhagium mordax

    AU - Friis, Dennis Steven

    AU - Johnsen, Johannes Lørup

    AU - Kristiansen, Erlend

    AU - Westh, Peter

    AU - Ramløv, Hans

    PY - 2014/6

    Y1 - 2014/6

    N2 - The equilibrium heat stability and the kinetic heat tolerance of a recombinant antifreeze protein (AFP) from the beetle Rhagium mordax (RmAFP1) are studied through differential scanning calorimetry and circular dichroism spectroscopy. In contrast to other insect AFPs studied with this respect, the RmAFP1 has only one disulfide bridge. The melting temperature, Tm, of the protein is determined to be 28.5°C (pH 7.4), which is much lower than most of those reported for AFPs or globular proteins in general. Despite its low melting temperature, both biophysical and activity measurements show that the protein almost completely refolds into the native state after repeated exposure of 70°C. RmAFP1 thus appears to be kinetically stable even far above its melting temperature. Thermodynamically, the insect AFPs seem to be dividable in three groups, relating to their content of disulfide bridges and widths of the ice binding motifs; high melting temperature AFPs (high disulfide content, TxT motifs), low melting temperature but high refolding capability AFPs (one disulfide bridge, TxTxTxT motifs) and irreversibly unfolded AFPs at low temperatures (no disulfide bridges, TxTxTxTxT motifs). The property of being able to cope with high temperature exposures may appear peculiar for proteins which strictly have their effect at subzero temperatures. Different aspects of this are discussed.

    AB - The equilibrium heat stability and the kinetic heat tolerance of a recombinant antifreeze protein (AFP) from the beetle Rhagium mordax (RmAFP1) are studied through differential scanning calorimetry and circular dichroism spectroscopy. In contrast to other insect AFPs studied with this respect, the RmAFP1 has only one disulfide bridge. The melting temperature, Tm, of the protein is determined to be 28.5°C (pH 7.4), which is much lower than most of those reported for AFPs or globular proteins in general. Despite its low melting temperature, both biophysical and activity measurements show that the protein almost completely refolds into the native state after repeated exposure of 70°C. RmAFP1 thus appears to be kinetically stable even far above its melting temperature. Thermodynamically, the insect AFPs seem to be dividable in three groups, relating to their content of disulfide bridges and widths of the ice binding motifs; high melting temperature AFPs (high disulfide content, TxT motifs), low melting temperature but high refolding capability AFPs (one disulfide bridge, TxTxTxT motifs) and irreversibly unfolded AFPs at low temperatures (no disulfide bridges, TxTxTxTxT motifs). The property of being able to cope with high temperature exposures may appear peculiar for proteins which strictly have their effect at subzero temperatures. Different aspects of this are discussed.

    KW - Antifreeze protein

    KW - Rhagium mordax

    KW - Stability

    KW - thermodynamic stability

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