Purification and characterization of a thermal hysteresis protein from a plant, the bittersweet nightshade Solanum dulcamara.

Thermal hysteresis proteins (THPs), which depress the freezing point of water below the melting point (producing a characteristic thermal hysteresis), are well known for their antifreeze activity in both fish and terrestrial arthropods, but have only recently been identified in plants. This study describes the purification of a THP from winter-collected bittersweet nightshade, Solanum dulcamara, using ion exchange and preparative 'free flow' isoelectric focusing. The THP has a molecular mass of 67 kDa (considerably larger than those of animal THPs), and an unusually high glycine component (23.7 mol%). Treatments of the THP with periodate or borate caused inactivation, suggesting the presence of carbohydrate. More specific treatments directed at galactose (beta-galactosidase or Abrus precatorius lectin) also resulted in inactivation, indicating that galactose is present. A thermal hysteresis activity versus THP concentration curve showed that the specific activity of the S. dulcamara THP is lower than that of any known animal THP. The functional significance of this low activity is discussed.