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Fluorescence microscopy evidence for quasi-permanent attachment of antifreeze proteins to ice surfaces
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Pertaya N Marshall CB DiPrinzio CL Wilen L Thomson ES Wettlaufer JS Davies PL Braslavsky I 《Biophysical journal》2007,92(10):3663-3673
Many organisms are protected from freezing by the presence of extracellular antifreeze proteins (AFPs), which bind to ice, modify its morphology, and prevent its further growth. These proteins have a wide range of applications including cryopreservation, frost protection, and as models in biomineralization research. However, understanding their mechanism of action remains an outstanding challenge. While the prevailing adsorption-inhibition hypothesis argues that AFPs must bind irreversibly to ice to arrest its growth, other theories suggest that there is exchange between the bound surface proteins and the free proteins in solution. By conjugating green fluorescence protein (GFP) to a fish AFP (Type III), we observed the binding of the AFP to ice. This was accomplished by monitoring the presence of GFP-AFP on the surface of ice crystals several microns in diameter using fluorescence microscopy. The lack of recovery of fluorescence after photobleaching of the GFP component of the surface-bound GFP-AFP shows that there is no equilibrium surface-solution exchange of GFP-AFP and thus supports the adsorption-inhibition mechanism for this type of AFP. Moreover, our study establishes the utility of fluorescently labeled AFPs as a research tool for investigating the mechanisms underlying the activity of this diverse group of proteins. 相似文献
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Neal G. Ravindra Mia Madel Alfajaro Victor Gasque Nicholas C. Huston Han Wan Klara Szigeti-Buck Yuki Yasumoto Allison M. Greaney Victoria Habet Ryan D. Chow Jennifer S. Chen Jin Wei Renata B. Filler Bao Wang Guilin Wang Laura E. Niklason Ruth R. Montgomery Stephanie C. Eisenbarth Sidi Chen Adam Williams Akiko Iwasaki Tamas L. Horvath Ellen F. Foxman Richard W. Pierce Anna Marie Pyle David van Dijk Craig B. Wilen 《PLoS biology》2021,19(3)
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