Direct visualization of spruce budworm antifreeze protein interacting with ice crystals: Basal plane affinity confers hyperactivity

Natalya Pertaya, Christopher B. Marshall, Yeliz Celik, Peter L. Davies, Ido Braslavsky*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

99 Scopus citations


Antifreeze proteins (AFPs) protect certain organisms from freezing by adhering to ice crystals, thereby preventing their growth. All AFPs depress the nonequilibrium freezing temperature below the melting point; however AFPs from overwintering insects, such as the spruce budworm (sbw) are 10-100 times more effective than most fish AFPs. It has been proposed that the exceptional activity of these AFPs depends on their ability to prevent ice growth at the basal plane. To test the hypothesis that the hyperactivity of sbwAFP results from direct affinity to the basal plane, we fluorescently tagged sbwAFP and visualized it on the surface of ice crystals using fluorescence microscopy. SbwAFP accumulated at the six prism plane corners and the two basal planes of hexagonal ice crystals. In contrast, fluorescently tagged fish type III AFP did not adhere to the basal planes of a single-crystal ice hemisphere. When ice crystals were grown in the presence of a mixture of type III AFP and sbwAFP, a hybrid crystal shape was produced with sbwAFP bound to the basal planes of truncated bipyramidal crystals. These observations are consistent with the blockage of c-axial growth of ice as a result of direct interaction of sbwAFP with the basal planes.

Original languageAmerican English
Pages (from-to)333-341
Number of pages9
JournalBiophysical Journal
Issue number1
StatePublished - 1 Jul 2008
Externally publishedYes

Bibliographical note

Funding Information:
This work was supported by the Canadian Institutes for Health Research to P.L.D who holds a Canada Research Chair in Protein Engineering, the Condensed Matter and Surface Science program at Ohio University and the Biomimetic Nanoscience and Nanoscale Technology initiative at Ohio University to I.B.


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