Identification and characterisation of novel inhibitors on extrinsic tenase complex from Bungarus fasciatus (banded krait) venom

Wan Chen; Leng Chuan Goh; Tse Siang Kang; Manjunatha R. Kini

doi:10.1160/TH13-12-1063

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2014; 112(04): 700-715
DOI: 10.1160/TH13-12-1063

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Identification and characterisation of novel inhibitors on extrinsic tenase complex from Bungarus fasciatus (banded krait) venom

Wan Chen^#

¹Department of Pharmacy, National University of Singapore, Singapore, Singapore

,

Leng Chuan Goh^#

¹Department of Pharmacy, National University of Singapore, Singapore, Singapore

,

Tse Siang Kang

¹Department of Pharmacy, National University of Singapore, Singapore, Singapore

,

Manjunatha R. Kini

²Department of Biological Sciences, National University of Singapore, Singapore, Singapore

³Department of Biochemistry and Molecular Biology, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia, USA

⁴School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, South Australia, Australia

› Institutsangaben

Abstract

Summary

Snake venoms are excellent sources of pharmacologically active proteins and peptides, and hence are potential sources of leads for drug developments. It has been previously established that krait (Bungarus genus) venoms contain mainly neurotoxins. A screening for anticoagulants showed that Bungarus fasciatus venom exhibits potent anticoagulant effect in standard clotting assays. Through sequential fractionation of the venom by size exclusion and high performance liquid chromatographies, coupled with functional screening for anticoagulant activities, we have isolated and purified two anticoagulant proteins, termed BF-AC1 ( Bungarus fasciatus anticoagulant 1) and BFAC2. They have potent inhibitory activities (IC₅₀ of 10 nM) on the extrinsic tenase complex. Structurally, these proteins each has two subunits covalently held together by disulfide bond(s). The N-terminal sequences of the individual subunits of BF-AC1 and BF-AC2 showed that the larger subunit is homologous to phospholipase A2, while the smaller subunit is homologous to Kunitz type serine proteinase inhibitor. Functionally, in addition to their anticoagulant activity, these proteins showed presynaptic neurotoxic effects in both in vivo and ex vivo experiments. Thus, BF-AC1 and BF-AC2 are structurally and functionally similar to β-bungarotoxins, a class of neurotoxins. The enzymatic activity of phospholipase A2 subunit plays a significant role in the anticoagulant activities. This is the first report on the anticoagulant activity of β-bungarotoxins and these results expand on the existing catalogue of haemostatically active snake venom proteins.

Keywords

Anticoagulant - extrinsic tenase complex - β-bungarotoxin - phospholipase A₂ - Bungarus fasciatus

Volltext

Referenzen

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