Purification and Characterization of a Novel Metalloproteinase, Acurhagin, from Agkistrodon acutus Venom

Wen-Jeng Wang; Tur-Fu Huang

doi:10.1055/s-0037-1613061

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2002; 87(04): 641-650
DOI: 10.1055/s-0037-1613061

Review Article

Schattauer GmbH

Purification and Characterization of a Novel Metalloproteinase, Acurhagin, from Agkistrodon acutus Venom

Wen-Jeng Wang

¹Department of Pharmacology, College of Medicine, National Taiwan University, Taipei, Taiwan

,

Tur-Fu Huang

¹Department of Pharmacology, College of Medicine, National Taiwan University, Taipei, Taiwan

› Author Affiliations

Abstract

Summary

Acurhagin, a high-molecular mass hemorrhagic metalloproteinase, was purified from the crude venom of Agkistrodon acutus using anionexchange and hydrophobic interaction chromatography. Acurhagin is a monomer with a molecular mass of 51.4 kDa under non-reducing conditions on SDS-PAGE and 48,133 Da by mass spectrometry. Partial amino acid sequence of its metalloproteinase domain is homologous to other high-molecular mass metalloproteinases from snake venoms. It preferentially cleaved Aα. chain of fibrinogen, followed by Bβ chain, while γ chains was minimally affected. Monitored by RP-HPLC, it extensively degraded fibrinogen into various peptide fragments. In aqueous solution, acurhagin autoproteolyzed to a 30 kDa fragment at 37° C. The N-terminal sequence of the 30 kDa fragment of acurhagin showed a high homology to those proteins consisting of disintegrinlike and cysteine-rich domains. Caseinolytic assay showed that the proteinase activity of acurhagin was slightly enhanced by Ca²⁺ and Mg²⁺, but completely inhibited by Zn²⁺. When treated with metal chelators, acurhagin was completely inactivated. Furthermore, acurhagin exerts an inhibitory effect on ADP-induced platelet aggregation of plateletrich plasma in an incubation-time dependent manner. It also impairs collagen- and ristocetin-induced platelet aggregation by cleaving collagen and vWF, respectively.

Keywords

Snake venom metalloproteinase - Agkistrodon acutus, fibrinogenase - autoproteolysis - platelet aggregation

Full Text

References

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