LmrTX, a basic PLA(2) (D49) purified from Lachesis muta rhombeata snake venom with enzymatic-related antithrombotic and anticoagulant activity

Author(s):Show less -

Damico, Daniela C. S. ^[1] ; Vassequi-Silva, T. ^[1] ; Torres-Huaco, F. D. ^[1] ; Nery-Diez, A. C. C. ^[1] ; de Souza, R. C. G. ^[2] ; Da Silva, S. L. ^[3] ; Vicente, C. P. ^[4] ; Mendes, C. B. ^[5] ; Antunes, E. ^[5] ; Werneck, C. C. ^[1] ; Marangoni, Sergio ^[1]

Affiliation

^[1] Univ Estadual Campinas, Dept Biochem, Inst Biol, UNICAMP, BR-13083970 Campinas, SP - Brazil

^[2] Hosp Fdn Itacare, Itacare, BA - Brazil

^[3] Fed Univ Sao Joao Del Rei UFSJ, Chem Biotecnol & Bioproc Dept, Ouro Branco, MG - Brazil

^[4] Univ Estadual Campinas, Dept Anat Cell Biol & Physiol & Biophys, Inst Biol, UNICAMP, BR-13083970 Campinas, SP - Brazil

^[5] Univ Estadual Campinas, Dept Pharmacol, Campinas, SP - Brazil

• Document type: Journal article
• Source: Toxicon; v. 60, n. 5, p. 773-781, OCT 2012.
• Web of Science Citations: 13

Abstract

A basic phospholipase A(2) (LmrTX) isoform was isolated from Lachesis muta rhombeata snake venom and partially characterized. The venom was fractionated by molecular exclusion chromatography in ammonium bicarbonate buffer followed by reverse-phase HPLC on a C-5 Discovery (R) Bio Wide column. From liquid chromatography-electrospray ionization/mass spectrometry, the molecular mass of LmrTX was measured as 14.277.50 Da. The amino acid sequence showed a high degree of homology between PLA(2) LmrTX from L muta rhombeata and other PLA(2) from snake venoms, like CB1 and CB2 from Crotalus durissus terrificus; LmTX-I and LmTX-II from Lachesis muta muta. LmrTX had PLA(2) activity in the presence of a synthetic substrate and alkylation of histidine residues significantly inhibited (P < 0.05) the enzymatic activity of LmrTX and its anticoagulant and antithrombotic activity. In this study, we examined the ability of the LmrTX in altering thrombus formation in living mouse, using a photochemically induced arterial thrombosis model. The control animals that did not receive protein injection showed a normal occlusion time, which was around 57 +/- 7.8 min. LmrTX, the PLA(2) from L. muta rhombeata venom, caused a change in the occlusion time to 99 +/- 10 min with doses of 7.5 mu g/mice. Additionally, LmrTX showed the anticoagulant activity in vitro and ex vivo and prolonging the time aggregation in wash platelet induced by ADP and Thrombin. (C) 2012 Elsevier Ltd. All rights reserved. (AU)

• FAPESP's process: 10/19916-7 - Evaluation on elastic fiber synthesis in cell culture obtained from fibrillin-1 defficient mice: Losartan's effect studies
• Grantee: Claudio Chrysostomo Werneck
• Support Opportunities: Regular Research Grants
• FAPESP's process: 09/02299-8 - Structure-function study of Lachesis muta rhombeata venom toxin active in thrombus formation in vivo
• Grantee: Daniela Carla da Silva Damico
• Support Opportunities: Scholarships in Brazil - Post-Doctoral