Isolation and structural characterization of a cytotoxic L-amino acid oxidase from Agkistrodon contortrix laticinctus snake venom: Preliminary crystallographic data

Citation
Dhf. Souza et al., Isolation and structural characterization of a cytotoxic L-amino acid oxidase from Agkistrodon contortrix laticinctus snake venom: Preliminary crystallographic data, ARCH BIOCH, 368(2), 1999, pp. 285-290
Citations number
32
Categorie Soggetti
Biochemistry & Biophysics
Journal title
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
ISSN journal
00039861 → ACNP
Volume
368
Issue
2
Year of publication
1999
Pages
285 - 290
Database
ISI
SICI code
0003-9861(19990815)368:2<285:IASCOA>2.0.ZU;2-Q
Abstract
We have purified a cytotoxic L-amino acid oxidase (LAO) from Agkistrodon co ntortrix laticinctus snake venom by means of Superdex-200 gel filtration, f ollowed 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 c ulture assay. After 24 h treatment with 25 mu g/ml of ACL LAO, the typical DNA fragmentation pattern of apoptotic cells was observed on agarose gel el ectrophoresis. NMR analysis showed the presence of a flavin mononucleotide prosthetic group. To solve its 3-D structure, crystals of the purified prot ein were grown in 0.1 M Tris-HCl, pH 8.5, and 2 M (NH4)(2)SO4. Diffraction data collected to 3.5 Angstrom showed that the protein crystallized in the tetragonal system, with unit cell a = b = 103.22 Angstrom, c = 183.45 Angst rom. This is the first report of preliminary crystallization data for a sna ke venom L-amino acid oxidase. (C) 1999 Academic Press.