Antifreeze Glycoproteins Inhibit Leakage from Liposomes during Thermotropic Phase Transitions

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 93; no. 13; pp. 6835 - 6840
• Main Authors: Hays, Lisa M., Feeney, Robert E., Crowe, Lois M., Crowe, John H., Oliver, Ann E.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 25.06.1996; National Acad Sciences; National Academy of Sciences
• Subjects: Animals; Antifreeze Proteins; Calorimetry, Differential Scanning; Cell Membrane Permeability - physiology; Cellular biology; Cooling; Fishes; Fluorescence; Freezing; Gels; Glycoproteins - isolation & purification; Glycoproteins - physiology; Lipids; Liposomes; Liquids; Marine; Membranes; Membranes, Artificial; Nototheniidae; Phase transitions; Phospholipids - physiology; Proteins; Temperature; Transition temperature; PS, Antarctica
• DOI: 10.1073/pnas.93.13.6835
• ISSN: 0027-8424

Antifreeze glycoproteins (AFGPs), found in the blood of polar fish at concentrations as high as 35 g/liter, are known to prevent ice crystal growth and depress the freezing temperature of the blood. Previously, Rubinsky et al. [Rubinsky, B., Mattioli, M., Arav, A., Barboni, B. & Fletcher, G. L. (1992) Am. J. Physiol. 262, R542-R545] provided evidence that AFGPs block ion fluxes across membranes during cooling, an effect that they ascribed to interactions with ion channels. We investigated the effects of AFGPs on the leakage of a trapped marker from liposomes during chilling. As these liposomes are cooled through the transition temperature, they leak ≈ 50% of their contents. Addition of less than 1 mg/ml of AFGP prevents up to 100% of this leakage, both during chilling and warming through the phase transition. This is a general effect that we show here applies to liposomes composed of phospholipids with transition temperatures ranging from 12 degrees C to 41 degrees C. Because these results were obtained with liposomes composed of phospholipids alone, we conclude that the stabilizing effects of AFGPs on intact cells during chilling reported by Rubinsky et al. may be due to a nonspecific effect on the lipid components of native membranes. There are other proteins that prevent leakage, but only under specialized conditions. For instance, antifreeze proteins, bovine serum albumin, and ovomucoid all either have no effect or actually induce leakage. Following precipitation with acetone, all three proteins inhibited leakage, although not to the extent seen with AFGPs. Alternatively, there are proteins such as ovotransferrin that have no effect on leakage, either before or after acetone precipitation.