Purification of antifreeze proteins by adsorption to ice.
Antifreeze proteins (AFPs) can protect organisms from freezing injury by adsorbing to ice and inhibiting its growth. We describe here a method where ice, grown on a cold finger, is used to selectively adsorb and purify these ice-binding proteins from a crude mixture. Type III recombinant AFP was enriched approximately 50-fold after one round of partitioning into ice and purified to homogeneity by a second round. This method can also be used to purify non-ice-binding proteins by linkage to AFP domains as demonstrated by the recovery of a 50 kDa maltose-binding protein-AFP fusion from a crude lysate of Escherichia coli.
Kuiper
Michael J
MJ
Department of Biology, Queen's University, Kingston, Ont., Canada K7L 3N6.
Lankin
Christopher
C
Gauthier
Sherry Y
SY
Walker
Virginia K
VK
Davies
Peter L
PL
D000327:Adsorption; D000818:Animals; D021343:Antifreeze Proteins, Type III / Q000737:chemistry* / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D005615:Freezing; D007053:Ice / Q000032:analysis; D012034:Refrigeration / Q000295:instrumentation / Q000379:methods
D021343:Antifreeze Proteins, Type III; D007053:Ice