Purification and characterization of two platelet-aggregation inhibitors, named angustatin and H-toxin TA(2), from the venom of Dendroaspis angusticeps

Etsuko Oyama; Hidenobu Takahashi

doi:10.1016/j.toxicon.2014.11.002

Purification and characterization of two platelet-aggregation inhibitors, named angustatin and H-toxin TA(2), from the venom of Dendroaspis angusticeps

Toxicon. 2015 Jan:93:61-7. doi: 10.1016/j.toxicon.2014.11.002. Epub 2014 Nov 5.

Authors

Etsuko Oyama¹, Hidenobu Takahashi²

Affiliations

¹ Department of Hygienic Chemistry, Meiji Pharmaceutical University, 2-522-1, Noshio, Kiyose, Tokyo 204-8588, Japan. Electronic address: oyama@my-pharm.ac.jp.
² Emeritus Professor of Meiji Pharmaceutical University, 2-522-1, Noshio, Kiyose, Tokyo 204-8588, Japan.

PMID: 25447773
DOI: 10.1016/j.toxicon.2014.11.002

Abstract

Angustatin and H-toxin TA2 were purified from unfractionated Dendroaspis angusticeps venom (0.3 g) using S-Sepharose fast flow column chromatography, gelfiltration on a Sephadex G-50 column, and reverse-phase HPLC. The purified materials strongly inhibited ADP-induced platelet aggregation. The primary structure of angustatin was determined by the Edman degradation of peptides derived from digestions with endopeptidese Arg-C and α-chymotrypsin. Angustatin, which was composed of 59 amino acid residues and an RGD sequence, was identified as a short-length inhibitor similar to mambin, dendroaspin, echistatin, and eristicophin. Angustatin shared 83%, 17%, and 15% homologies with mambin, eristicophin, and echistatin, respectively. On the other hand, H-toxin TA2 did not conserve the RGD sequence in its structure; it was replaced for the Glu-Met-Leu sequence. Furthermore, the amino acid sequence of H-toxin TA2 corresponded to toxin TA2, excluding the amino acid residue of His28Arg.

Keywords: Inhibitor; Neurotoxin; Platelet aggregation; Snake venom.

MeSH terms

Amino Acid Sequence
Animals
Base Sequence
Chromatography, High Pressure Liquid
Elapid Venoms / analysis
Elapid Venoms / chemistry*
Elapid Venoms / isolation & purification
Elapidae*
Molecular Sequence Data
Platelet Aggregation Inhibitors / analysis
Platelet Aggregation Inhibitors / isolation & purification*
Sequence Analysis, DNA

Substances

Elapid Venoms
H-toxin TA2, Dendroaspis angusticeps
Platelet Aggregation Inhibitors
angustatin, Dendroaspis angusticeps