Inhibition of bacterial ice nucleators by fish antifreeze glycoproteins

Abstract

Certain bacteria promote the formation of ice in super-cooled water by means of ice nucleators which contain a unique protein associated with the cell membrane¹. Ice nucleators in general are believed to act by mimicking the structure of an ice crystal surface, thus imposing an ice-like arrangement on the water molecules in contact with the nucleating surface and lowering the energy necessary for the initiation of ice formation². Quantitative investi-gation of the bacterial ice-nucleating process has recently been made possible by the discovery of certain bacteria that shed stable membrane vesicles with ice nucleating activity³. The opposite effect, inhibition of ice formation, has been described for a group of glycoproteins found in different fish and insect species^4,5. This group of substances, termed antifreeze glycoproteins (AFGPs), promotes the supercooling of water with no appreciable effect on the equilibrium freezing point or melting temperature⁶. Substantial evidence now indicates that AFGPs act by binding to a growing ice crystal and slowing crystal growth^7,8. As the ice-nucleating protein surface is believed to have a structure similar to an embry-onic ice crystal, AFGPs might be predicted to interact directly with a bacterial ice-nucleating site. We report here that AFGPs from the antarctic fish Dissostichus mamoni inhibit the ice-nucleating activity of membrane vesicles from the bacterium Erwinia herbicola. The inhibition effect shows saturation at high concentration of AFGP and conforms to a simple binding reaction between the AFGP and the nucleation centre.