Structure of an antifreeze polypeptide precursor from the sea raven, Hemitripterus americanus.
The cystine-rich antifreeze polypeptides (AFP) from sea raven were fractionated by reverse-phase high performance liquid chromatography into several components, with SR2 (Mr 17,000) as the major AFP. Sea raven AFP cDNA clones were isolated from a liver cDNA library using a synthetic oligonucleotide, and the identity of one of the clones, C2-1, was confirmed by hybridization selection and cell-free translation. C2-1 encodes a pre-AFP of 195 amino acids with no evidence of any profragments. Comparison of the deduced amino acid sequence with partial peptide sequences from SR2 showed substitutions in at least four amino acid positions, suggesting that C2-1 cDNA codes for a minor component. Both the primary and the predicted secondary structures of sea raven AFP are completely different from those of other fish AFP. This further confirms that sea raven AFP belongs to a different class of antifreezes. The high frequency of reverse turns and the presence of paired hydrophilic amino acids in these structures are striking features of the protein and may contribute to their antifreeze action.
D000595:Amino Acid Sequence; D000818:Animals; D021301:Antifreeze Proteins; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant; D005399:Fishes / Q000097:blood*; D006023:Glycoproteins / Q000097:blood* / Q000235:genetics; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D011487:Protein Conformation; D011498:Protein Precursors / Q000097:blood*
D021301:Antifreeze Proteins; D004274:DNA, Recombinant; D006023:Glycoproteins; D011498:Protein Precursors; D004247:DNA