Lufaxin, a novel factor Xa inhibitor from the salivary gland of the sand fly Lutzomyia longipalpis blocks protease-activated receptor 2 activation and inhibits inflammation and thrombosis in vivo.

Détails

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Etat: Serval
Version: de l'auteur
ID Serval
serval:BIB_1CBA6DC2B671
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Lufaxin, a novel factor Xa inhibitor from the salivary gland of the sand fly Lutzomyia longipalpis blocks protease-activated receptor 2 activation and inhibits inflammation and thrombosis in vivo.
Périodique
Arteriosclerosis, Thrombosis, and Vascular Biology
Auteur(s)
Collin N., Assumpção T.C., Mizurini D.M., Gilmore D.C., Dutra-Oliveira A., Kotsyfakis M., Sá-Nunes A., Teixeira C., Ribeiro J.M., Monteiro R.Q., Valenzuela J.G., Francischetti I.M.
ISSN
1524-4636 (Electronic)
ISSN-L
1079-5642
Statut éditorial
Publié
Date de publication
2012
Peer-reviewed
Oui
Volume
32
Numéro
9
Pages
2185-2198
Langue
anglais
Résumé
OBJECTIVE: Blood-sucking arthropods' salivary glands contain a remarkable diversity of antihemostatics. The aim of the present study was to identify the unique salivary anticoagulant of the sand fly Lutzomyia longipalpis, which remained elusive for decades.
METHODS AND RESULTS: Several L. longipalpis salivary proteins were expressed in human embryonic kidney 293 cells and screened for inhibition of blood coagulation. A novel 32.4-kDa molecule, named Lufaxin, was identified as a slow, tight, noncompetitive, and reversible inhibitor of factor Xa (FXa). Notably, Lufaxin's primary sequence does not share similarity to any physiological or salivary inhibitors of coagulation reported to date. Lufaxin is specific for FXa and does not interact with FX, Dansyl-Glu-Gly-Arg-FXa, or 15 other enzymes. In addition, Lufaxin blocks prothrombinase and increases both prothrombin time and activated partial thromboplastin time. Surface plasmon resonance experiments revealed that FXa binds Lufaxin with an equilibrium constant ≈3 nM, and isothermal titration calorimetry determined a stoichiometry of 1:1. Lufaxin also prevents protease-activated receptor 2 activation by FXa in the MDA-MB-231 cell line and abrogates edema formation triggered by injection of FXa in the paw of mice. Moreover, Lufaxin prevents FeCl(3)-induced carotid artery thrombus formation and prolongs activated partial thromboplastin time ex vivo, implying that it works as an anticoagulant in vivo. Finally, salivary gland of sand flies was found to inhibit FXa and to interact with the enzyme.
CONCLUSIONS: Lufaxin belongs to a novel family of slow-tight FXa inhibitors, which display antithrombotic and anti-inflammatory activities. It is a useful tool to understand FXa structural features and its role in prohemostatic and proinflammatory events.
Mots-clé
Amino Acid Sequence, Animals, Anti-Inflammatory Agents/chemistry, Anti-Inflammatory Agents/isolation & purification, Blood Coagulation/drug effects, Calorimetry, Cell Line, Tumor, Chlorides, Cloning, Molecular, Disease Models, Animal, Dose-Response Relationship, Drug, Factor Xa/antagonists & inhibitors, Factor Xa/metabolism, Female, Ferric Compounds, Fibrinolytic Agents/chemistry, Fibrinolytic Agents/isolation & purification, HEK293 Cells, Humans, Inflammation/blood, Inflammation/metabolism, Insect Proteins/chemistry, Insect Proteins/isolation & purification, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Molecular Weight, Partial Thromboplastin Time, Protein Binding, Prothrombin Time, Psychodidae/chemistry, Rats, Receptor, PAR-2/antagonists & inhibitors, Receptor, PAR-2/metabolism, Recombinant Proteins/antagonists & inhibitors, Recombinant Proteins/metabolism, Salivary Glands/chemistry, Surface Plasmon Resonance, Thromboplastin/antagonists & inhibitors, Thromboplastin/metabolism, Thrombosis/blood, Thrombosis/chemically induced, Time Factors
Pubmed
Web of science
Création de la notice
17/09/2012 14:53
Dernière modification de la notice
03/03/2018 14:32
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