Antifreeze peptide heterogeneity in an antarctic eel pout includes an unusually large major variant comprised of two 7 kDa type III AFPs linked in tandem.
The structural heterogeneity of the major antifreeze peptides (AFPs) from the antarctic eel pout, Lycodichthys dearborni (formerly classified as Rhigophila dearborni) was characterized. Three major AFPs designated as RD1, RD2 and RD3, and five minor ones were isolated from the fish plasma. RD1 and RD2 are both 64 residues in length, about 7 kDa, and thus similar in size to all characterized type III AFPs, while RD3 is twice as large, about 14 kDa, and represents the first example of a disparately large size variant within the same fish for the three known types of antifreeze peptides. RD3 was found to be 134 residues in length, arranged as a 64-residue N-terminal half and a 61-residue C-terminal half of similar sequence to each other and to the 7 kDa type III AFPs, linked by a 9-residue connector of unmatched sequence. RD3 has slightly lower antifreeze activity than its 7 kDa counterparts, with a melting-freezing point difference of about 0.81 degrees C at 10 mg/ml versus 0.95 degrees C and 0.90 degrees C for RD1 and RD2, respectively. RD1 and RD2 are 94% identical in sequence to each other. They are 98% and 94%, respectively identical to N-terminal half of RD3, and 85% and 77%, respectively, identical to C-terminal half of RD3. By sequence comparison, a previously characterized AFP from this fish [1] was identified to be RD2.
Wang
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Department of Physiology, University of Illinois, Urbana 61801.
DeVries
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Cheng
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D000595:Amino Acid Sequence; D000818:Animals; D000864:Antarctic Regions; D021301:Antifreeze Proteins; D021343:Antifreeze Proteins, Type III; D004524:Eels / Q000097:blood*; D010450:Endopeptidases; D005615:Freezing; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D016415:Sequence Alignment; D014357:Trypsin
D021301:Antifreeze Proteins; D021343:Antifreeze Proteins, Type III; D006023:Glycoproteins; D011506:Proteins; C092813:RD3 protein, eel; D010450:Endopeptidases; D014357:Trypsin