Structural and functional properties of Bp-LAAO, a new L-amino acid oxidase isolated from Bothrops pauloensis snake venom.
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.
Rodrigues
Renata S
RS
Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, UFU, Pará 1720, CEP 38400-902 Uberlândia, MG, Brazil.
da Silva
Juliana F
JF
Boldrini França
Johara
J
Fonseca
Fernando P P
FP
Otaviano
Antônio R
AR
Henrique Silva
Flávio
F
Hamaguchi
Amélia
A
Magro
Angelo J
AJ
Braz
Antônio Sérgio K
AS
dos Santos
Juliana I
JI
Homsi-Brandeburgo
Maria Inês
MI
Fontes
Marcos R M
MR
Fuly
André L
AL
Soares
Andreimar M
AM
Rodrigues
Veridiana M
VM
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
D003435:Crotalid Venoms; D050830:L-Amino Acid Oxidase