A New Factor Xa Inhibitor (Lefaxin) from the Haementeria depressa Leech

Fernanda Faria; Eva Maria A. Kelen; Claudio A. M. Sampaio; Cassian Bon; Nathalie Duval; Ana Marisa Chudzinski-Tavassi

doi:10.1055/s-0037-1614857

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1999; 82(05): 1469-1473
DOI: 10.1055/s-0037-1614857

Rapid Communications

Schattauer GmbH

A New Factor Xa Inhibitor (Lefaxin) from the Haementeria depressa Leech

Fernanda Faria

¹From Laboratorio de Fisiopatologia, Instituto Butantan; São-Paulo, Brasil

,

Eva Maria A. Kelen

¹From Laboratorio de Fisiopatologia, Instituto Butantan; São-Paulo, Brasil

,

Claudio A. M. Sampaio

²Departamento de Bioquímica e Biologia Molecular, Universidade Federal de São Paulo (UNIFESP-EPM) São Paulo, Brasil

,

Cassian Bon

³Unité des Venins, Institut Pasteur, Paris, France

,

Nathalie Duval^*

³Unité des Venins, Institut Pasteur, Paris, France

,

Ana Marisa Chudzinski-Tavassi

¹From Laboratorio de Fisiopatologia, Instituto Butantan; São-Paulo, Brasil

› Author Affiliations

Abstract

Summary

The salivary complex of the leech Haementeria depressa produces potent anticoagulant components. Among them, a protein named lefaxin inhibits factor Xa (FXa). Lefaxin was purified to homogeneity from dissected salivary complexes by gel filtration in Sephadex G-150 followed by two ion exchange chromatography steps in Mono-Q. Inhibition of FXa by lefaxin was demonstrated by the inhibition of its amidolytic activity, measured with chromogenic substrate S-2765 (apparent K_I of 4 nM), and of its ability to inhibit thrombin generation in the prothrombinase complex (EC₅₀ of 40 nM). Lefaxin has a molecular weight of 30 kDa and an isoelectric point of 5.7. It is made of a polypeptide chain whose N-terminal sequence shows no similarity with that of other FXa inhibitors (antistasin and ghilianten) isolated from leech saliva. On the other hand, the N-terminal sequence of lefaxin presents significant sequence similarity with nitric oxide carrier proteins myohemerythrin from the annelid Nereis diversicolor and prolixin S from the tick Rhodnius prolixus. Interestingly, prolixin S also proved to be an anticoagulant protein acting on FXa.

Keywords

Leeches - FXa inhibitor - Haementeria depressa

Full Text

References

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