title: Microfluidic experiments reveal that antifreeze proteins bound to ice crystals suffice to prevent their growth.
abstract: Antifreeze proteins (AFPs) are a subset of ice-binding proteins that control ice crystal growth. They have potential for the cryopreservation of cells, tissues, and organs, as well as for production and storage of food and protection of crops from frost. However, the detailed mechanism of action of AFPs is still unclear. Specifically, there is controversy regarding reversibility of binding of AFPs to crystal surfaces. The experimentally observed dependence of activity of AFPs on their concentration in solution appears to indicate that the binding is reversible. Here, by a series of experiments in temperature-controlled microfluidic devices, where the medium surrounding ice crystals can be exchanged, we show that the binding of hyperactive Tenebrio molitor AFP to ice crystals is practically irreversible and that surface-bound AFPs are sufficient to inhibit ice crystal growth even in solutions depleted of AFPs. These findings rule out theories of AFP activity relying on the presence of unbound protein molecules.
authors.affiliation: Department of Physics and Astronomy, Ohio University, Athens, OH 45701, USA.
authors.forename: Yeliz
authors.initials: Y
authors.lastname: Celik
authors.forename: Ran
authors.initials: R
authors.lastname: Drori
authors.forename: Natalya
authors.initials: N
authors.lastname: Pertaya-Braun
authors.forename: Aysun
authors.initials: A
authors.lastname: Altan
authors.forename: Tyler
authors.initials: T
authors.lastname: Barton
authors.forename: Maya
authors.initials: M
authors.lastname: Bar-Dolev
authors.forename: Alex
authors.initials: A
authors.lastname: Groisman
authors.forename: Peter L
authors.initials: PL
authors.lastname: Davies
authors.forename: Ido
authors.initials: I
authors.lastname: Braslavsky
mesh_terms: D000818:Animals; D021301:Antifreeze Proteins / Q000737:chemistry* / Q000378:metabolism*; D055592:Biophysical Phenomena; D007053:Ice; D019476:Insect Proteins / Q000737:chemistry / Q000378:metabolism; D046210:Microfluidic Analytical Techniques; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013712:Tenebrio / Q000378:metabolism
chemical_list: D021301:Antifreeze Proteins; D007053:Ice; D019476:Insect Proteins; D011993:Recombinant Fusion Proteins