Effect of Type I Antifreeze Proteins on the Freezing and Melting Processes of Cryoprotective Solutions Studied by Site-Directed Spin Labeling Technique

Antifreeze proteins (AFPs) protect organisms living in subzero environments from freezing injury, which render them potential applications for cryopreservation of living cells, organs, and tissues. Cryoprotective agents (CPAs), such as glycerol and propylene glycol, have been used as ingredients to...

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Published inCrystals (Basel) Vol. 9; no. 7; p. 352
Main Authors Perez, Adiel F., Taing, Kyle R., Quon, Justin C., Flores, Antonia, Ba, Yong
Format Journal Article
LanguageEnglish
Published Basel MDPI AG 01.07.2019
Subjects
antifreeze protein
Cooling
cryo-photo microscopy
Cryopreservation
cryoprotective agent
Electron paramagnetic resonance
Freezing
Glycerol
Hemodialysis
ice crystal
Ice crystals
Ice formation
Labeling
Low temperature
Melting points
Nitrogen
Organs
Peptides
Propylene
Proteins
Room temperature
spin labeling
United States > US
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Summary:Antifreeze proteins (AFPs) protect organisms living in subzero environments from freezing injury, which render them potential applications for cryopreservation of living cells, organs, and tissues. Cryoprotective agents (CPAs), such as glycerol and propylene glycol, have been used as ingredients to treat cellular tissues and organs to prevent ice crystal’s formation at low temperatures. To assess AFP’s function in CPA solutions, we have the applied site-directed spin labeling technique to a Type I AFP. A two-step process to prevent bulk freezing of the CPA solutions was observed by the cryo-photo microscopy, i.e., (1) thermodynamic freezing point depression by the CPAs; and (2) inhibition to the growth of seed ice crystals by the AFP. Electron paramagnetic resonance (EPR) experiments were also carried out from room temperature to 97 K, and vice versa. The EPR results indicate that the spin labeled AFP bound to ice surfaces, and inhibit the growths of ice through the bulk freezing processes in the CPA solutions. The ice-surface bound AFP in the frozen matrices could also prevent the formation of large ice crystals during the melting processes of the solutions. Our study illustrates that AFPs can play an active role in CPA solutions for cryopreservation applications.
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Author Contributions: Conceptualization, Y.B.; methodology, Y.B., A.P., K.T., J.Q., and A.F.; software, Y.B., A.P., K.T., J.Q., and A.F.; validation, Y.B., A.P., K.T., J.Q.; formal analysis, Y.B., A.P., K.T., J.Q., and A.F.; investigation, Y.B., A.P., K.T., J.Q., and A.F.; resources, Y.B.; data curation, Y.B., A.P., K.T., J.Q., and A.F.; writing—original draft preparation, Y.B., A.P.; writing—review and editing, Y.B., A.P., K.T. and J.Q.; visualization, Y.B., A.P., K.T., J.Q., and A.F.; supervision, Y.B.; project administration, Y.B.; funding acquisition, Y.B.
ISSN:2073-4352
2073-4352
DOI:10.3390/cryst9070352