Document Detail


Biochemical, pharmacological and structural characterization of two PLA2 isoforms Cdr-12 and Cdr-13 from Crotalus durissus ruruima snake venom.
MedLine Citation:
PMID:  17203396     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cdr-12 and Cdr-13 isoforms of PLA2, a D49 protein, were purified from Crotalus durissus ruruima venom after one chromatographic step, reverse phase HPLC on micro-Bondapack C-18. The molecular mass by SDS-PAGE of Cdr-12 and Cdr-13 isoforms of PLA2 was 14333.49 Da and 14296.42 Da, respectively and confirmed by MALDI-TOF mass spectrometry. The amino acid composition showed that both isoforms Cdr-12 and Cdr-13 have a high content of Lys, Tyr, Gly, Arg, and 14 half-Cys residues, typical of a basic PLA2. The isoforms Cdr-12 and Cdr-13 had a sequence of amino acids of 122 amino acid residues, being Cdr-12: SLLQFNKMIK FETRKNAIPF YAFYGCYCGW GGQGRPKDAT DRCCIVHDCC YGKLAKCNTK WDFYRYSLRS GYFQCGKGTW CEQQICECDR VAAECLRRSL STYRYGYMIY PDSRCREPSE TC and pI value 8.37 and Cdr-13: SLVQFEKMIK EETGKNAVPF YAFYGCYCGW GGRGRPKDAT DRCCIVHDCC YEKLVKCNTK WDFYRYSLRS GYFQCGKGTW CEQQICECDR VAAECLRRSL STYRYGKMIY PDSRCREPSE TC with a pI value of 8.13 This sequence shows high identity values when compared to other D49 PLA2s isolated from venoms of crotalics snakes. Skeletal muscle preparations from the young chicken have been previously used in order to study the effects of toxins on neuromuscular transmission, providing an important opportunity to study the differentiated behavior of a toxin before more than one model, because it shows differences in its sensibilities. In mice, the PLA2 isoforms Cdr-12 and Cdr-13 induced myonecrosis and edema, upon intramuscular or subcutaneous injections, respectively. In vitro, Cdr-12 and Cdr-13 isoforms of PLA2, caused a potent blockade of neuromuscular transmission in young chicken biventer cervicis preparation and produced cytotoxicity in murine C2C12 skeletal muscle myotubes and lack cytolytic activity upon myoblasts in vitro. Thus, the combined structural and functional information obtained identify Cdr-12 and Cdr-13 isoforms as members of the PLA2 family, which presents the typical bioactivities described for such proteins.
Authors:
Luis Alberto Ponce-Soto; Paulo Aparecido Baldasso; Frey Francisco Romero-Vargas; Flávia V Winck; José Camillo Novello; Sergio Marangoni
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The protein journal     Volume:  26     ISSN:  1572-3887     ISO Abbreviation:  Protein J.     Publication Date:  2007 Jan 
Date Detail:
Created Date:  2007-07-04     Completed Date:  2007-10-11     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  101212092     Medline TA:  Protein J     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  39-49     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, Institute of Biology, State University of Campinas, P.O. Box 6109, Zip code 13083-970, Campinas, SP, Brazil. poncesoto@yahoo.com.ar
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Cells, Cultured
Chemical Fractionation
Chickens
Crotalid Venoms / chemistry,  enzymology*,  isolation & purification,  toxicity
Crotalus
Diaphragm / drug effects
Isoelectric Point
Isoenzymes
Mice
Molecular Weight
Muscle, Skeletal / pathology
Myoblasts / drug effects,  metabolism
Necrosis / pathology
Phospholipases A / chemistry*,  isolation & purification,  toxicity*
Phospholipases A2
Phrenic Nerve / drug effects
Sequence Alignment
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Chemical
Reg. No./Substance:
0/Crotalid Venoms; 0/Isoenzymes; EC 3.1.1.-/Phospholipases A; EC 3.1.1.4/Cdr-12 protein, Crotalus durissus ruruima; EC 3.1.1.4/Cdr-13 protein, Crotalus durissus ruruima; EC 3.1.1.4/Phospholipases A2

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