Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/197650
Type: Artigo de periódico
Title: Structural And Functional Properties Of Batx, A New Lys49 Phospholipase A2 Homologue Isolated From The Venom Of The Snake Bothrops Alternatus.
Author: Ponce-Soto, Luis Alberto
Lomonte, Bruno
Gutiérrez, José María
Rodrigues-Simioni, Lea
Novello, José Camillo
Marangoni, Sergio
Abstract: BaTX PLA(2), a K49 phospholipase A(2) homologue was purified from Bothrops alternatus venom after two chromatographic steps, molecular exclusion on Superdex 75 and reverse phase HPLC on mu-Bondapack C-18. A molecular mass of 13898.71 Da was determined by MALDI-TOF mass spectrometry. The amino acid composition showed that BaTX has a high content of Lys, Tyr, Gly, Pro, and 14 half-Cys residues, typical of a basic PLA(2). The complete amino acid sequence of BaTX PLA(2) contains 121 residues, resulting in a calculated pI value of 8.63. This sequence shows high identity values when compared to other K49 PLA(2)s isolated from the venoms of viperid snakes. Lower identity is observed in comparison to D49 PLA(2)s. The sequence was SLFELGKMIL QETGKNPAKS YGAYYCYCGW GGQGQPKDAT DRCCYVHKCC YKKLTGCNPK KDRYSYSWKD KTIVCGENNS CLKELCECDK AVAICLRENL NTYNKKYRYY LKPLCKKADA C. In mice, BaTX induced myonecrosis and edema, upon intramuscular or subcutaneous injections, respectively. The LD(50) of BaTX was 7 mug/g body weight, by intravenous route. In vitro, the toxin caused a potent blockade of neuromuscular transmission in young chicken biventer cervicis preparations. The blockage 50% was achieved at a concentration of 0.03 microM: 40+/-0.4 min and 0.07 microM: 35+/-0.3 min. Moreover, this protein induced a rapid cytolytic effect upon mouse skeletal muscle myoblasts in culture. Thus, the combined structural and functional information obtained identify BaTX as a new member of the K49 PLA(2) family, which presents the typical bioactivities described for such proteins.
Subject: Amino Acid Sequence
Animals
Bothrops
Cell Line
Cell Survival
Chickens
Chromatography, Gel
Chromatography, High Pressure Liquid
Crotalid Venoms
Dose-response Relationship, Drug
Edema
In Vitro Techniques
Isoenzymes
Lethal Dose 50
Lysine
Mice
Molecular Sequence Data
Molecular Weight
Muscle, Skeletal
Myoblasts, Skeletal
Necrosis
Neuromuscular Junction
Phospholipases A
Phospholipases A2
Protein Conformation
Sequence Analysis, Protein
Sequence Homology, Amino Acid
Spectrometry, Mass, Matrix-assisted Laser Desorption-ionization
Time Factors
Rights: fechado
Identifier DOI: 10.1016/j.bbagen.2006.11.015
Address: http://www.ncbi.nlm.nih.gov/pubmed/17270350
Date Issue: 2007
Appears in Collections:Unicamp - Artigos e Outros Documentos

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