Document Detail


Structural and functional properties of BaTX, a new Lys49 phospholipase A2 homologue isolated from the venom of the snake Bothrops alternatus.
MedLine Citation:
PMID:  17270350     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Luis Alberto Ponce-Soto; Bruno Lomonte; José María Gutiérrez; Lea Rodrigues-Simioni; José Camillo Novello; Sergio Marangoni
Publication Detail:
Type:  Comparative Study; In Vitro; Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-12-15
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  1770     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  2007 Apr 
Date Detail:
Created Date:  2007-02-26     Completed Date:  2007-04-17     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  585-93     Citation Subset:  IM    
Affiliation:
Biochemistry Department, Institute of Biology, Medical Sciences Faculty, State University of Campinas (UNICAMP), Campinas, SP, Brazil. poncesoto@yahoo.com.ar
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Bothrops / metabolism*
Cell Line
Cell Survival / drug effects
Chickens
Chromatography, Gel
Chromatography, High Pressure Liquid
Crotalid Venoms / enzymology*
Dose-Response Relationship, Drug
Edema / chemically induced
Isoenzymes / chemistry
Lethal Dose 50
Lysine
Mice
Molecular Sequence Data
Molecular Weight
Muscle, Skeletal / drug effects,  pathology
Myoblasts, Skeletal / drug effects
Necrosis
Neuromuscular Junction / drug effects
Phospholipases A / chemistry*,  isolation & purification,  toxicity*
Phospholipases A2
Protein Conformation
Sequence Analysis, Protein
Sequence Homology, Amino Acid
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Time Factors
Chemical
Reg. No./Substance:
0/Crotalid Venoms; 0/Isoenzymes; 56-87-1/Lysine; EC 3.1.1.-/Phospholipases A; EC 3.1.1.4/Phospholipases A2

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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