| Isolation and structural characterization of a cytotoxic L-amino acid oxidase from Agkistrodon contortrix laticinctus snake venom: preliminary crystallographic data. | |
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MedLine Citation:
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PMID: 10441379 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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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:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Archives of biochemistry and biophysics Volume: 368 ISSN: 0003-9861 ISO Abbreviation: Arch. Biochem. Biophys. Publication Date: 1999 Aug |
Date Detail:
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Created Date: 1999-09-09 Completed Date: 1999-09-09 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0372430 Medline TA: Arch Biochem Biophys Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 285-90 Citation Subset: IM |
Copyright Information:
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Copyright 1999 Academic Press. |
Affiliation:
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Departamento de Ciências Fisiológicas, Universidade Federal de São Carlos, São Carlos, SP, 13565-905, Brazil. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Agkistrodon Amino Acid Oxidoreductases / chemistry, isolation & purification*, toxicity Animals Apoptosis / drug effects Crotalid Venoms / enzymology* DNA Fragmentation / drug effects HL-60 Cells Humans L-Amino Acid Oxidase Protein Conformation |
| Chemical | |
Reg. No./Substance:
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0/Crotalid Venoms; EC 1.4.-/Amino Acid Oxidoreductases; 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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