title: Cytotoxic L-amino acid oxidase from Bothrops moojeni: biochemical and functional characterization.
abstract: An L-amino acid oxidase isolated from Bothrops moojeni snake venom (BmooLAAO-I) was purified to a high degree using sequential CM-Sepharose ion-exchange and phenyl-Sepharose chromatography. When analyzed by mass spectrometry, the purified BmooLAAO-I presented a molecular weight of 64,889 and 130,779 under denaturing and nondenaturing conditions, respectively. BmooLAAO-I is a homodimeric acidic glycoprotein with a pI approximately 4.7, and the N-terminal sequence shows close structural similarity to other snake venom LAAOs. This enzyme was inactivated by freezing or low pH, and secondary structural analysis by circular dichroism revealed 48% alpha-helix, 20% beta-sheet, 12% beta-turn, and 20% random coil structures. BmooLAAO-I exhibited bactericidal, antitumoral, trypanocidal, edematogenic, and platelet-aggregating activities. All of these effects were inhibited by catalase, suggesting that these biological effects are mediated by the production of H(2)O(2). BmooLAAO-I induced typical apoptotic DNA fragmentation in HL-60 cells, which was also inhibited by catalase. These results point to the potential use of BmooLAAO-I as a therapeutic agent for treatment of diseases in which induction of H(2)O(2) production can be beneficial.
authors.affiliation: Laboratório de Bioquímica do Instituto de Pesquisas em Patologias Tropicais (IPEPATRO), Universidade Federal de Rondônia, UNIR, Porto Velho-RO, Brazil.
authors.forename: Rodrigo G
authors.initials: RG
authors.lastname: Stábeli
authors.forename: Carolina D
authors.initials: CD
authors.lastname: Sant'Ana
authors.forename: Patrícia H
authors.initials: PH
authors.lastname: Ribeiro
authors.forename: Tássia R
authors.initials: TR
authors.lastname: Costa
authors.forename: Fábio K
authors.initials: FK
authors.lastname: Ticli
authors.forename: Matheus G
authors.initials: MG
authors.lastname: Pires
authors.forename: Auro
authors.initials: A
authors.lastname: Nomizo
authors.forename: Sérgio
authors.initials: S
authors.lastname: Albuquerque
authors.forename: Natael R
authors.initials: NR
authors.lastname: Malta-Neto
authors.forename: Mozart
authors.initials: M
authors.lastname: Marins
authors.forename: Suely V
authors.initials: SV
authors.lastname: Sampaio
authors.forename: Andreimar M
authors.initials: AM
authors.lastname: Soares
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D000818:Animals; D017837:Bothrops; D002374:Catalase / Q000494:pharmacology; D002853:Chromatography, Liquid; D003435:Crotalid Venoms / Q000201:enzymology*; D053938:DNA Fragmentation / Q000187:drug effects*; D018922:HL-60 Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D017433:Protein Structure, Secondary
chemical_list: D003435:Crotalid Venoms; D006861:Hydrogen Peroxide; D002374:Catalase; D000594:Amino Acid Oxidoreductases