Purification, properties and cDNA cloning of neoverrucotoxin (neoVTX), a hemolytic lethal factor from the stonefish Synanceia verrucosa venom.
A proteinaceous toxin with hemolytic and lethal activities, named neoverrucotoxin (neoVTX), was purified from the venom fluid of stonefish Synanceia verrucosa and its primary structure was elucidated by a cDNA cloning technique. NeoVTX is a dimeric 166 kDa protein composed of alpha-subunit (702 amino acid residues) and beta-subunit (699 amino acid residues) and lacks carbohydrate moieties. Its hemolytic activity is inhibited by anionic lipids, especially potently by cardiolipin. These properties are comparable to those of stonustoxin (SNTX) previously purified from S. horrida. Alignment of the amino acid sequences also reveals that the neoVTX alpha- and beta-subunits share as high as 87 and 95% sequence identity with the SNTX alpha- and beta-subunits, respectively. The distinct differences between neoVTX and SNTX are recognized only in the numbers of Cys residues (18 for neoVTX and 15 for SNTX) and free thiol groups (10 for neoVTX and 5 for SNTX). In contrast, neoVTX considerably differs from verrucotoxin (VTX), a tetrameric 322 kDa glycoprotein, previously purified from S. verrucosa. In addition, the sequence identity of the neoVTX beta-subunit with the reported VTX beta-subunit is 90%, being lower than that with the SNTX beta-subunit.
Ueda
Atsushi
A
Department of Food Science and Technology, Tokyo University of Marine Science and Technology, Konan-4, Tokyo 108-8477, Japan.
Suzuki
Mika
M
Honma
Tomohiro
T
Nagai
Hiroshi
H
Nagashima
Yuji
Y
Shiomi
Kazuo
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D029941:Fish Proteins / Q000235:genetics* / Q000302:isolation & purification; D005397:Fish Venoms / Q000235:genetics* / Q000302:isolation & purification; D005400:Fishes, Poisonous / Q000235:genetics*; D008969:Molecular Sequence Data; D021122:Protein Subunits / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D029941:Fish Proteins; D005397:Fish Venoms; D021122:Protein Subunits