The amino acid sequence of antistasin. A potent inhibitor of factor Xa reveals a repeated internal structure.
Antistasin is a 15-kDa protein from the salivary glands of the Mexican leech, Haementeria officinalis, which manifests anticoagulant activity by inhibiting factor Xa. Previous work demonstrating the presence of this activity in salivary gland extracts and its partial purification has been reported (Tuszynski, G. P., Gasic, T. B, and Gasic, G.J. (1987) J. Biol. Chem. 262, 9718-9723). The present study includes further purification to homogeneity of antistasin and its subsequent fragmentation and complete amino acid sequence determination. The protein, which possesses 119 amino acid residues, is blocked at its amino terminus by the presence of a pyroglutamic acid residue and has an unusually high cysteine content, with 20 cysteine residues. The primary structure of antistasin shows no homology to hirudin, a 65-residue anticoagulant protein from the medicinal leech, Hirudo medicinalis. Of great interest is the finding of significant internal homology within antistasin where a 2-fold internal repeated structure is observed. At least four isoforms of antistasin have been identified in leech salivary gland extracts by high performance liquid chromatography analysis, and partial amino acid sequence analysis of these isoforms indicates they differ by 1 or 2 amino acid residues.
Nutt
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E
Merck Sharp & Dohme Research Laboratories, West Point, Pennsylvania 19486.
Gasic
T
T
Rodkey
J
J
Gasic
G J
GJ
Jacobs
J W
JW
Friedman
P A
PA
Simpson
E
E
D000595:Amino Acid Sequence; D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D015951:Factor Xa; D007447:Invertebrate Hormones / Q000302:isolation & purification / Q000494:pharmacology; D007865:Leeches; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D012469:Salivary Glands / Q000032:analysis; D015842:Serine Proteinase Inhibitors
D000925:Anticoagulants; D007447:Invertebrate Hormones; D010446:Peptide Fragments; D015842:Serine Proteinase Inhibitors; C053155:antistasin; D015951:Factor Xa