Isolation and characterization of antifreeze proteins from smelt (Osmerus mordax) and Atlantic herring (Clupea harengus harengus)

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Abstract
Antifreeze proteins (AFPs) from smelt (Osmerus mordax) and Atlantic herring (Clupea harengus harengus) were isolated using gel filtration, ion exchange chromatography, and high performance liquid chromatography. The AFPs of smelt appeared to consist of at least six components and those of Atlantic herring, of at least two components. The relative molecular masses of these antifreezes were 24 000 and 14 600, respectively. Amino acid analysis showed both proteins to be cystine-rich, type II AFPs like those of the sea raven (Hemitripterus americanus). In addition, smelt AFPs were found to be immunologically similar to those of the sea raven. The smelt AFPs differed from those of Atlantic herring and sea raven in that they contained a small amount of glucosamine (~3%). The activity levels of the smelt and herring AFPs were reduced in the presence of dithiothreitol, indicating the functional importance of intact disulfide bonds.