Document Detail

Isolation and structural characterization of a cytotoxic L-amino acid oxidase from Agkistrodon contortrix laticinctus snake venom: preliminary crystallographic data.
MedLine Citation:
PMID:  10441379     Owner:  NLM     Status:  MEDLINE    
We have purified a cytotoxic L-amino acid oxidase (LAO) from Agkistrodon contortrix laticinctus snake venom by means of Superdex-200 gel filtration, followed by phenyl-Sepharose CL-4B chromatography. The purified enzyme (ACL LAO) is a dimer on gel filtration, with a M(r) of 60,000 for the monomer as estimated by SDS-PAGE. LAO activity was tested against 15 amino acids, but only 9 were oxidized by the enzyme, suggesting that it presents some degree of specificity. ACL LAO has apoptosis-inducing activity in an HL-60 cell culture assay. After 24 h treatment with 25 micrograms/ml of ACL LAO, the typical DNA fragmentation pattern of apoptotic cells was observed on agarose gel electrophoresis. NMR analysis showed the presence of a flavin mononucleotide prosthetic group. To solve its 3-D structure, crystals of the purified protein were grown in 0.1 M Tris-HCl, pH 8.5, and 2 M (NH(4))(2)SO(4). Diffraction data collected to 3.5 A showed that the protein crystallized in the tetragonal system, with unit cell a = b = 103.22 A, c = 183.45 A. This is the first report of preliminary crystallization data for a snake venom L-amino acid oxidase.
D H Souza; L M Eugenio; J E Fletcher; M S Jiang; R C Garratt; G Oliva; H S Selistre-de-Araujo
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Archives of biochemistry and biophysics     Volume:  368     ISSN:  0003-9861     ISO Abbreviation:  Arch. Biochem. Biophys.     Publication Date:  1999 Aug 
Date Detail:
Created Date:  1999-09-09     Completed Date:  1999-09-09     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0372430     Medline TA:  Arch Biochem Biophys     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  285-90     Citation Subset:  IM    
Copyright Information:
Copyright 1999 Academic Press.
Departamento de Ciências Fisiológicas, Universidade Federal de São Carlos, São Carlos, SP, 13565-905, Brazil.
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MeSH Terms
Amino Acid Oxidoreductases / chemistry,  isolation & purification*,  toxicity
Apoptosis / drug effects
Crotalid Venoms / enzymology*
DNA Fragmentation / drug effects
HL-60 Cells
L-Amino Acid Oxidase
Protein Conformation
Reg. No./Substance:
0/Crotalid Venoms; EC 1.4.-/Amino Acid Oxidoreductases; EC Acid Oxidase

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