Adsorption Kinetics of Type-III Antifreeze Protein on Ice Crystal Surfaces

Abstract

The spatio-temporal distribution of type-III antifreeze protein (AFP-III) molecules labeled with fluorescent isocyanate (FITC) was visualized at the interfaces between ice and solutions with an FITC-labeled AFP-III (F-AFP-III) concentration of 20−800 μg/mL by fluorescence microscopy. The number density of F-AFP-III on the surface of ice microcrystals was calculated from the calibrated fluorescence intensity. The adsorption of F-AFP-III molecules on the ice crystal surfaces proceeded at a finite rate and then reached the saturation level. The time course of the number density of adsorbed F-AFP-III molecules could be well represented by Langmuir's model. The characteristic adsorption time of F-AFP-III, the adsorption coefficient k 1 = (0.5 ± 0.05) × 10 −4 (μg/mL) −1 s −1 , and the desorption coefficient k 2 = 0.005 ± 0.002 s −1 were determined using the Langmuir's model and obtained experimental data. We found that the adsorption of F-AFP-III could have different kinetics depending on the solution conditions and the type of fluorescence molecules conjugated with AFP-III.