Structural variations in the alanine-rich antifreeze proteins of the pleuronectinae.
The sequence and activity of antifreeze proteins from two right eye flounder species were compared to assess the influence of structural variations on antifreeze capacity. The cDNA encoding the major serum antifreeze protein in the yellowtail flounder (Limanda ferruginea) was cloned from liver tissue. Its DNA sequence shows that the precursor to the antifreeze is a 97-residue preproportion. Edman degradation identified the N-terminus of the 48-amino-acid mature serum antifreeze protein and confirmed the sequence of the first 36 residues. A comparison with the previously determined winter flounder antifreeze protein and mRNA sequences shows strong homology through the 5' and 3' untranslated regions and in the peptide region. The mature protein section has the greatest sequence variation. Specifically, the yellowtail antifreeze protein, in contrast to that of the winter flounder, contains a fourth 11-amino-acid repeat and lacks several of the hydrophilic residues that have been postulated to aid in the binding of the protein to ice crystals. Intramolecular salt bridges are present in the antifreeze proteins from both species but in different registries with respect to the 11-amino-acid repeats. On a mass basis the yellowtail flounder antifreeze, though longer than that of the winter flounder, is only 80% as effective at depressing the freezing temperature of aqueous solutions. This lower activity might be due to the reduced number of hydrophilic ice-binding residues per molecule.
Scott
G K
GK
Department of Biochemistry, Queen's University, Kingston, Ontario, Canada.
Davies
P L
PL
Shears
M A
MA
Fletcher
G L
GL
D000409:Alanine / Q000032:analysis*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D021301:Antifreeze Proteins; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D005412:Flatfishes / Q000378:metabolism*; D005432:Flounder / Q000378:metabolism*; D006023:Glycoproteins / Q000032:analysis* / Q000235:genetics; D008099:Liver / Q000378:metabolism; D008961:Models, Structural; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000032:analysis; D013696:Temperature
D000596:Amino Acids; D021301:Antifreeze Proteins; D006023:Glycoproteins; D011498:Protein Precursors; D004247:DNA; D000409:Alanine