Dendrimer-Linked Antifreeze Proteins Have Superior Activity and Thermal Recovery

Bioconjug Chem. 2015 Sep 16;26(9):1908-15. doi: 10.1021/acs.bioconjchem.5b00290. Epub 2015 Aug 20.

Abstract

By binding to ice, antifreeze proteins (AFPs) depress the freezing point of a solution and inhibit ice recrystallization if freezing does occur. Previous work showed that the activity of an AFP was incrementally increased by fusing it to another protein. Even larger increases in activity were achieved by doubling the number of ice-binding sites by dimerization. Here, we have combined the two strategies by linking multiple outward-facing AFPs to a dendrimer to significantly increase both the size of the molecule and the number of ice-binding sites. Using a heterobifunctional cross-linker, we attached between 6 and 11 type III AFPs to a second-generation polyamidoamine (G2-PAMAM) dendrimer with 16 reactive termini. This heterogeneous sample of dendrimer-linked type III constructs showed a greater than 4-fold increase in freezing point depression over that of monomeric type III AFP. This multimerized AFP was particularly effective at ice recrystallization inhibition activity, likely because it can simultaneously bind multiple ice surfaces. Additionally, attachment to the dendrimer has afforded the AFP superior recovery from heat denaturation. Linking AFPs together via polymers can generate novel reagents for controlling ice growth and recrystallization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antifreeze Proteins / chemistry*
  • Crystallization
  • Dendrimers / chemistry*
  • Freezing*
  • Ice
  • Models, Molecular
  • Molecular Weight
  • Protein Conformation

Substances

  • Antifreeze Proteins
  • Dendrimers
  • Ice
  • PAMAM Starburst