title: Structural and functional properties of Bp-LAAO, a new L-amino acid oxidase isolated from Bothrops pauloensis snake venom.
abstract: An L-amino acid oxidase (Bp-LAAO) from Bothrops pauloensis snake venom was highly purified using sequential chromatography steps on CM-Sepharose, Phenyl-Sepharose CL-4B, Benzamidine Sepharose and C18 reverse-phase HPLC. Purified Bp-LAAO showed to be a homodimeric acidic glycoprotein with molecular weight around 65kDa under reducing conditions in SDS-PAGE. The best substrates for Bp-LAAO were L-Met, L-Leu, L-Phe and L-Ile and the enzyme showed a strong reduction of its catalytic activity upon L-Met and L-Phe substrates at extreme temperatures. Bp-LAAO showed leishmanicidal, antitumoral and bactericidal activities dose dependently. Bp-LAAO induced platelet aggregation in platelet-rich plasma and this activity was inhibited by catalase. Bp-LAAO-cDNA of 1548bp codified a mature protein with 516 amino acid residues corresponding to a theoretical isoelectric point and molecular weight of 6.3 and 58kDa, respectively. Additionally, structural and phylogenetic studies identified residues under positive selection and their probable location in Bp-LAAO and other snake venom LAAOs (svLAAOs). Structural and functional investigations of these enzymes can contribute to the advancement of toxinology and to the elaboration of novel therapeutic agents.
authors.affiliation: Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, UFU, Pará 1720, CEP 38400-902 Uberlândia, MG, Brazil.
authors.forename: Renata S
authors.initials: RS
authors.lastname: Rodrigues
authors.forename: Juliana F
authors.initials: JF
authors.lastname: da Silva
authors.forename: Johara
authors.initials: J
authors.lastname: Boldrini França
authors.forename: Fernando P P
authors.initials: FP
authors.lastname: Fonseca
authors.forename: Antônio R
authors.initials: AR
authors.lastname: Otaviano
authors.forename: Flávio
authors.initials: F
authors.lastname: Henrique Silva
authors.forename: Amélia
authors.initials: A
authors.lastname: Hamaguchi
authors.forename: Angelo J
authors.initials: AJ
authors.lastname: Magro
authors.forename: Antônio Sérgio K
authors.initials: AS
authors.lastname: Braz
authors.forename: Juliana I
authors.initials: JI
authors.lastname: dos Santos
authors.forename: Maria Inês
authors.initials: MI
authors.lastname: Homsi-Brandeburgo
authors.forename: Marcos R M
authors.initials: MR
authors.lastname: Fontes
authors.forename: André L
authors.initials: AL
authors.lastname: Fuly
authors.forename: Andreimar M
authors.initials: AM
authors.lastname: Soares
authors.forename: Veridiana M
authors.initials: VM
authors.lastname: Rodrigues
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017837:Bothrops / Q000378:metabolism*; D001943:Breast Neoplasms / Q000378:metabolism; D045744:Cell Line, Tumor; D003435:Crotalid Venoms / Q000201:enzymology*; D004926:Escherichia coli / Q000187:drug effects; D006801:Humans; D050830:L-Amino Acid Oxidase / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D007891:Leishmania / Q000187:drug effects; D015458:Leukemia, T-Cell / Q000378:metabolism; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010802:Phylogeny; D010974:Platelet Aggregation / Q000187:drug effects; D016415:Sequence Alignment; D013211:Staphylococcus aureus / Q000187:drug effects; D013379:Substrate Specificity / Q000502:physiology
chemical_list: D003435:Crotalid Venoms; D050830:L-Amino Acid Oxidase