Gene duplication of coagulation factor V and origin of venom prothrombin activator in Pseudonaja textilis snake

Thi Nguyet Minh Le; Md Abu Reza; Sanjay Swarup; R. Manjunatha Kini

doi:10.1160/TH04-11-0707

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2005; 93(03): 420-429
DOI: 10.1160/TH04-11-0707

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Gene duplication of coagulation factor V and origin of venom prothrombin activator in Pseudonaja textilis snake

Thi Nguyet Minh Le

¹Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, Republic of Singapore

,

Md Abu Reza

¹Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, Republic of Singapore

,

Sanjay Swarup

¹Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, Republic of Singapore

,

R. Manjunatha Kini

¹Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, Republic of Singapore

²Department of Biochemistry and Molecular Biophysics, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia, USA

› Author Affiliations

Abstract

Summary

The origin and evolution of venom toxins is a mystery that has evoked much interest. We have recently shown that pseutarin C, a prothrombin activator from Pseudonaja textilis venom, is structurally and functionally similar to mammalian coagulation factor Xa – factor Va complex. Its catalytic subunit is homologous to factor Xa while the nonenzymatic subunit is homologous to factor Va. P.textilis therefore has two parallel prothrombin activator systems: one expressed in its venom gland as a toxin and the other expressed in its liver and released into its plasma as a haemostatic factor. Here we report the complete amino acid sequence of factor V (FV) from its liver determined by cDNA cloning and sequencing. The liver FV shows 96% identity to pseutarin C nonenzymatic subunit. Most of the functional sites involved in its interaction with factor Xa and prothrombin are conserved. However, many potential sites of post-translational modifications and one critical cleavage site for activated protein C are different. The absence of the latter cleavage site makes pseutarin C nonenzymatic subunit resistant to inactivation and enhances its potential as an excellent toxin. By PCR and real-time quantitative analysis, we show that pseutarin C nonenzymatic subunit gene is expressed specifically in the venom gland at ~280 fold higher than that of FV gene in liver. These two are thus encoded by two separate genes that express in a highly tissue-specific manner. Our results imply that the gene encoding pseutarin C nonenzymatic subunit was derived by the duplication of plasma FV gene and they have evolved to perform distinct functions.

Keywords

Coagulation factor V - gene duplication - Pseudonaja textilis - venom toxin evolution

Full Text

References

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