Letter

β-Helix structure and ice-binding properties of a hyperactive antifreeze protein from an insect

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Abstract

Insect antifreeze proteins (AFP) are considerably more active at inhibiting ice crystal growth than AFP from fish or plants. Several insect AFPs, also known as thermal hysteresis proteins, have been cloned1,2,3 and expressed1,2. Their maximum activity is 3–4 times that of fish AFPs1 and they are 10–100 times more effective at micromolar concentrations. Here we report the solution structure of spruce budworm (Choristoneura fumiferana) AFP and characterize its ice-binding properties. The 9-kDa AFP is a β-helix with a triangular cross-section and rectangular sides that form stacked parallel β-sheets; a fold which is distinct from the three known fish AFP structures. The ice-binding side contains 9 of the 14 surface-accessible threonines organized in a regular array of TXT motifs that match the ice lattice on both prism and basal planes. In support of this model, ice crystal morphology and ice-etching experiments are consistent with AFP binding to both of these planes and thus may explain the greater activity of the spruce budworm antifreeze.

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Acknowledgements

We thank S. Gauthier and D. Doucet for help with the mutagenesis, and L. Saltibus and G. McQuaid for their excellent technical assistance. Special thanks to L. Spyracopoulos for helping in the analysis of the NMR data and structures. We thank Dan Garrett and the Laboratory of Chemical Physics at the National Institutes of Health for making available the program PIPP that was used in analysing our NMR data. This work was supported by MRC grants to P.L.D,B.D.S and Z.J. and an NSERC grant to V.K.W.; Z.J. is an MRC Scholar and P.L.D. is a Killam Research Fellow.

Author information

Author notes

    • Steffen P. Graether

    Present address: Department of Biochemistry, University of Alberta, Edmonton, Alberta T6G 2H7 , Canada.

Affiliations

  1. *Department of Biochemistry, Queen's University, Kingston, Ontario K7L 3N6, Canada

    • Steffen P. Graether
    • , Zongchao Jia
    •  & Peter L. Davies
  2. ‡Department of Biology, Queen's University , Kingston, Ontario K7L 3N6, Canada

    • Michael J. Kuiper
    • , Virginia K. Walker
    •  & Peter L. Davies
  3. §Department of Biochemistry, University of Alberta, Edmonton, Alberta T6G 2H7, Canada

    • Stéphane M. Gagné
    •  & Brian D. Sykes

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Corresponding author

Correspondence to Peter L. Davies.

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