Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/196965
Type: Artigo de periódico
Title: Cytotoxic Action In Myoblasts And Myotubes (c2c12) And Enzymatic Characterization Of A New Phospholipase A2 Isoform (bj-v) From Bothrops Jararacussu Venom.
Author: Bonfim, Vera Luis
Ponce-Soto, Luis Alberto
Novello, Jose Camilo
Marangoni, Sergio
Abstract: A new PLA2 Bj-V from Bothrops jararacussu (14039.49 Da determined by MALDI-TOF mass spectrometry) was isolated in only one chromatographic step by HPLC ion-exchange and its purity was confirmed by reverse phase. Amino acid analysis showed a high content of hydrophobic and basic amino acids as well as 14 half-cysteine residues. The N-terminal sequence (DLWQFGQMIL KETGKIPFPY YGAYGCYCGW GGRGGKPKDG TDRCCYVHD...) showed a high degree of homology with basic D49 PLA2 myotoxins from other Bothrops venoms. Bj V showed discrete sigmoidal enzymatic behavior, with maximal activity at pH 8.4 and 35-40 degrees C. Full PLA2 activity required Ca2+ (10 mM) and there was little catalytic activity in the presence of 1 mM Ca2+. The addition of Mn2+ or Mg2+ (10 mM) in the presence of low (1 mM) Ca2+ slightly increased the enzyme activity, whereas Zn2+ and Cu2+ (10 mM) diminished the activity. The substitution of Ca2+ for Mg2+ or Cu2+ also reduced the enzymatic activity. Bj V had PLA2 activity and produced cytotoxicity in murine C2C12 skeletal muscle myoblasts and myotubes. The isolation of these isoforms Bj-IV [1] and Bj-V (described herein) found in a fraction previously described as homogeneous shows us the importance of optimization in purification techniques in order to better understand their biological behavior.
Subject: Amino Acid Sequence
Animals
Bothrops
Cell Line
Crotalid Venoms
Isoenzymes
Mice
Molecular Sequence Data
Muscle Fibers, Skeletal
Myoblasts
Phospholipases A
Phospholipases A2
Rights: fechado
Identifier DOI: 
Address: http://www.ncbi.nlm.nih.gov/pubmed/17018014
Date Issue: 2006
Appears in Collections:Unicamp - Artigos e Outros Documentos

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