Cytotoxic l-amino acid oxidase from Bothrops moojeni: Biochemical and functional characterization

• Published in: International journal of biological macromolecules Vol. 41; no. 2; pp. 132 - 140
• Main Authors: Stábeli, Rodrigo G., Sant’Ana, Carolina D., Ribeiro, Patrícia H., Costa, Tássia R., Ticli, Fábio K., Pires, Matheus G., Nomizo, Auro, Albuquerque, Sérgio, Malta-Neto, Natael R., Marins, Mozart, Sampaio, Suely V., Soares, Andreimar M.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.07.2007
• Subjects: Amino Acid Oxidoreductases - chemistry; Amino Acid Oxidoreductases - isolation & purification; Amino Acid Oxidoreductases - pharmacology; Animals; Antimicrobial and cytotoxic effects; Bothrops; Bothrops moojeni; Catalase - pharmacology; Chromatography, Liquid; Crotalid Venoms - enzymology; DNA Fragmentation - drug effects; HL-60 Cells; Humans; Hydrogen Peroxide - metabolism; l-Amino acid oxidase; Platelet aggregation; Protein Structure, Secondary; Snake venom; Structure analysis; LAAO; l-Amino acid oxidase; BmooLAAO-I; Snake venom; Antimicrobial and cytotoxic effects; Bothrops moojeni; Platelet aggregation; sv-LAAO; Structure analysis
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2007.01.006

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 p I ∼ 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% α-helix, 20% β-sheet, 12% β-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 2O 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 2O 2 production can be beneficial.