Artificial multimers of the type III antifreeze protein. Effects on thermal hysteresis and ice crystal morphology.
A variant of antifreeze protein (AFP) named RD3 from antarctic eel pout (Lycodichthys dearborni) comprises the type III AFP intramolecular dimer, which is known to exhibit a significant enhancement of thermal hysteresis when compared with the type III AFP monomer (Miura, K., Ohgiya, S., Hoshino, T, Nemoto, N., Suetake, T., Miura, A, Spyracopoulos, L., Kondo, H., and Tsuda, S. (2001) J. Biol. Chem. 276, 1304-1310). Here we genetically synthesized intramolecular dimer, trimer, and tetramer of the type III AFP, for which we utilize the genes encoding the primary sequences of the N-domain, the C-domain, and the 9-residue linker of RD3, and we examined the AFP multimerization effects on thermal hysteresis and ice crystal morphology. Significantly, (i) the thermal hysteresis increases in proportion with the size of the multimers, (ii) a larger size of the multimer exerts the maximum activity at lower concentration, (iii) every multimer changes the morphology of a single ice crystal into a unique shape that is similar but not identical to the ordinary hexagonal bipyramid, and (iv) the size of ice crystal becomes dramatically small with increasing the concentration of the multimer. The thermal hysteresis enhancement of the multimer was detected in both molar and domain bases. These results suggest that a molecule comprising the multiple AFP domains connected in tandem acquires an enhanced affinity for the ice binding.
Nishimiya
Yoshiyuki
Y
Protein Structure Research Group, Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, 2-17-2-1 Tsukisamu-Higashi, Toyohira, Sapporo 062-8517, Japan.
Ohgiya
Satoru
S
Tsuda
Sakae
S
D021343:Antifreeze Proteins, Type III / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D003460:Crystallization; D017931:DNA Primers; D007053:Ice; D008958:Models, Molecular; D011487:Protein Conformation
D021343:Antifreeze Proteins, Type III; D017931:DNA Primers; D007053:Ice