F-segments of Arabidopsis dehydrins show cryoprotective activities for lactate dehydrogenase depending on the hydrophobic residues.

Although dehydrins show cryoprotective activities for freeze-sensitive enzymes, the underlying mechanism is still under investigation. Here, we report that F-segments conserved in some dehydrins cryoprotected lactate dehydrogenase (LDH) as well as K-segments, which were previously identified as cryoprotective segments of dehydrins. The cryoprotective activity levels of four F-segments of Arabidopsis dehydrins were similar to that of a typical K-segment. Amino acid substitution experiments indicated that the activity of the F-segment of Arabidopsis COR47 (designated as Fseg) depended on the hydrophobic residues (L, F, and V). Intriguingly, when all the amino acids other than the hydrophobic residues were changed to glycine, the cryoprotective activity did not change, suggesting that the hydrophobic amino acids were sufficient for Fseg activity. Circular dichroism analysis indicated that Fseg was mainly disordered in aqueous solution as well as Fseg_Φ/T, in which the hydrophobic residues of Fseg were changed to T. This suggested that the hydrophobic interaction might be related to the cryoprotective activities of Fseg.