Document Detail


Isolation of a new L-amino acid oxidase from Crotalus durissus cascavella venom.
MedLine Citation:
PMID:  16307769     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A novel l-amino acid oxidase (LAO) (Casca LAO) from Crotalus durissus cascavella venom was purified to a high degree of molecular homogeneity using a combination of molecular exclusion and ion-exchange chromatography system. The purified monomer of LAO presented a molecular mass of 68 kDa and pI estimated in 5.43, which were determined by two-dimensional electrophoresis. The 71st N-terminal amino acid sequence of the LAO from Crotalus durissus cascavella presented a high amino acid sequence similarities with other LAOs from Colloselasma rhosostoma, Crotalus adamanteus, Agkistrodon h. blomhoffi, Agkistrodon h. halys and Trimeresurus stejnegeri. LAO displayed a Michaelis-Menten behavior with a kilometer of 46.7 microM and an optimum pH for enzymatic activity of 6.5. Casca LAO induced a dose-dependent platelet aggregation, which was abolished by catalase and inhibited by indomethacin and aspirin. These results suggest that the production of H2O2 is involved in subsequent activation of inflammatory enzymes, such as thromboxane. Casca LAO also inhibited the bacterial growth of Gram-negative (Xanthomonas axonopodis pv passiflorae) and Gram-positive (S. mutans) strains. Electron microscopy assessments of both bacterial strains suggest that the hydrogen peroxide produced by LAO induce bacterial membrane rupture and consequently loss of cytoplasmatic content. This LAO exhibited a high antileishmanic activity against the promastigote of Leishmania amazonensis in vitro, its activity was dependent on the production of hydrogen peroxide, and the 50% inhibitory concentration was estimated in 2.39 microg/ml.
Authors:
Marcos H Toyama; Daniela de O Toyama; Luiz F D Passero; Marcia D Laurenti; Carlos E Corbett; Thaise Y Tomokane; Fabiana V Fonseca; Edson Antunes; Paulo P Joazeiro; Luis O S Beriam; Maria Alice C Martins; Helena S A Monteiro; Marasses C Fonteles
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2005-11-22
Journal Detail:
Title:  Toxicon : official journal of the International Society on Toxinology     Volume:  47     ISSN:  0041-0101     ISO Abbreviation:  Toxicon     Publication Date:  2006 Jan 
Date Detail:
Created Date:  2006-01-09     Completed Date:  2006-04-17     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  1307333     Medline TA:  Toxicon     Country:  England    
Other Details:
Languages:  eng     Pagination:  47-57     Citation Subset:  IM    
Affiliation:
UNESP, Campus do Litoral Paulista, São Vicente, São Paulo, São Paulo, Brazil. mhtjpn@yahoo.com
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Anti-Bacterial Agents / pharmacology
Bacteria / drug effects*
Crotalid Venoms / enzymology*
Dose-Response Relationship, Drug
Hydrogen Peroxide / metabolism
Hydrogen-Ion Concentration
L-Amino Acid Oxidase / isolation & purification,  pharmacology*
Leishmaniasis / drug therapy
Molecular Sequence Data
Platelet Aggregation / drug effects*
Platelet Aggregation Inhibitors / pharmacology
Time Factors
Chemical
Reg. No./Substance:
0/Anti-Bacterial Agents; 0/Crotalid Venoms; 0/Platelet Aggregation Inhibitors; 7722-84-1/Hydrogen Peroxide; EC 1.4.3.2/L-Amino Acid Oxidase

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


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