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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 |
Citation: | Protein And Peptide Letters. v. 13, n. 7, p. 707-13, 2006. |
Rights: | fechado |
Address: | http://www.ncbi.nlm.nih.gov/pubmed/17018014 |
Date Issue: | 2006 |
Appears in Collections: | Unicamp - Artigos e Outros Documentos |
Files in This Item:
File | Size | Format | |
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pmed_17018014.pdf | 300.34 kB | Adobe PDF | View/Open |
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