Thermal hysteresis of two antifreeze proteins from Fragilariopsis cylindrus (Bacillariophyceae)

Antifreeze proteins (AFPs) provide protection for organisms subjected to the presence of ice crystals. The psychrophilic diatom Fragilariopsis cylindrus which is frequently found in polar sea ice carries a multitude of AFP isoforms. In this study we report the heterologous expression of two antifreeze protein isoforms from F. cylindrus in Escherichia coli. Refolding from inclusion bodies produced proteins functionally active with respect to crystal deformation, recrystallization inhibition and thermal hysteresis. We observed a reduction of activity in the presence of the pelB leader peptide in comparison with the GS-linked SUMO-tag. Activity was positively correlated to protein concentration and buffer salinity. Thermal hysteresis and crystal deformation habit suggest the affiliation of the proteins to the hyperactive group of AFPs. One isoform, carrying a signal peptide for secretion, produced a thermal hysteresis up to 1.53 °C ± 0.53 °C and ice crystals of hexagonal bipyramidal shape. The second isoform, which has a long preceding N-terminal sequence of unknown function, produced thermal hysteresis of up to 2.34 °C ± 0.25 °C. Ice crystals grew in form of a hexagonal column in presence of this protein. The different sequences preceding the ice binding domain point to distinct localizations of the proteins inside or outside the cell. We thus propose that AFPs have different functions in vivo, also reflected in their specific TH capability.

Composition of the argillite sequence, continental slope of the Sea of Japan

The argillite sequence located at the base of the sedimentary cover on the continental slope of the Sea of Japan was studied by petrographic, palynological, and X-ray diffraction methods. Two spores-pollen complexes were distinguished in it: the Late Oligocene reflecting cooling and the Early Miocene corresponding to initiated warming. Data obtained indicate that the sequence is composed of terrigenous silty-clayey sediments that accumulated in shallow coastal-marine settings. The global sea-level rise at the Early-Middle Miocene transition, combined with regional tectonic processes, determined basin's deepening, owing to which the argillite sequence was overlain by a thick layer of Middle Miocene diatomaceous-clayey sediments. Due to tectonic movement along existing faults in the terminal Late Miocene, the argillite sequence occurring initially at depths of at least 400-500 m was locally exhumed to the basin bottom.