Isolation and characterization of an antifreeze protein from the longhorn sculpin, Myoxocephalus octodecimspinosis.
A new type of antifreeze protein was isolated from the serum of the longhorn sculpin, Myoxocephalus octodecimspinosis, by gel filtration and high-performance liquid chromatography. This protein (LS-12) exhibits freezing point depression activity (thermal hysteresis) and ice crystal modification properties similar to those seen for other types of fish antifreeze polypeptide, except that ice crystals grow as hexagonal trapezohedra in the presence of LS-12, rather than hexagonal bipyramids usually seen. Ice crystal etching studies demonstrate that LS-12 does not bind to the hexagonal bipyramidal or secondary prism surfaces reported for the antifreeze polypeptides from winter flounder and shorthorn sculpin, respectively. Circular dichroism studies indicate that LS-12 has an alpha-helix content of about 60% at 1 degreesC, which is in good agreement with a value of about 70% predicted from the amino acid sequence. Limited proteolysis studies and further analysis of the amino acid sequence suggest that LS-12 consists of four amphipathic alpha-helices of similar length which are folded into a four-helix bundle. Based on its size (Mr=12299) and predicted tertiary structure, LS-12 can be regarded as the first example of a new class (type IV) of fish antifreeze protein.
Deng
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Department of Chemistry, Boston University, Boston, MA 02215, USA.
Laursen
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RA
D000595:Amino Acid Sequence; D021301:Antifreeze Proteins; D021302:Antifreeze Proteins, Type IV; D002942:Circular Dichroism; D003461:Crystallography; D029941:Fish Proteins; D005615:Freezing; D006023:Glycoproteins / Q000737:chemistry*; D007053:Ice; D008969:Molecular Sequence Data; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D013696:Temperature; D013820:Thermolysin / Q000378:metabolism
D021301:Antifreeze Proteins; D021302:Antifreeze Proteins, Type IV; D029941:Fish Proteins; D006023:Glycoproteins; D007053:Ice; C103901:LS-12 antifreeze protein, Myoxocephalus octodecemspinosus; D011506:Proteins; D013820:Thermolysin