Biochemical and functional characterization of an L-amino acid oxidase isolated from Bothrops pirajai snake venom.
In this work we describe the isolation of a new l-amino acid oxidase (LAAO) referred to as BpirLAAO-I from Bothrops pirajai snake venom, which was highly purified using a combination of molecular exclusion, affinity, and hydrophobic chromatography steps. BpirLAAO-I homodimeric acid glycoprotein (approximate Mr and pI of 130,000 and 4.9, respectively) displays high specificity toward hydrophobic/aromatic amino acids, while deglycosylation does not alter its enzymatic activity. The N-terminal LAAO sequence of its first 49 amino acids presented a high similarity between a amino acid sequence with other LAAOs from: Bothrops spp., Crotalus spp., Calloselasma rhodostoma, Agkistrodon spp., Trimeresurus spp., Pseudechis australis, Oxyuranus scutellatus, and Notechis scutatus. BpirLAAO-I induces time-dependent platelet aggregation, mouse paw edema, cytotoxic activity against Escherichia coli, Pseudomonas aeruginosa, Leishmania sp., and tumor cells, and also a typical fago (M13mp18) DNA fragmentation. Platelet aggregation, leishmanicidal and antitumoral activities were reduced by catalase. Thus, BpirLAAO-I is a multifunctional protein with promising biotechnological and medical applications.
Izidoro
Luiz Fernando M
LF
Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, UFU, 38400-902 Uberlândia-MG, Brazil.
Ribeiro
Marcela C
MC
Souza
Guilherme R L
GR
Sant'Ana
Carolina D
CD
Hamaguchi
Amelia
A
Homsi-Brandeburgo
Maria I
MI
Goulart
Luiz R
LR
Beleboni
Rene O
RO
Nomizo
Auro
A
Sampaio
Suely V
SV
Soares
Andreimar M
AM
Rodrigues
Veridiana M
VM
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D017837:Bothrops; D002384:Catalysis; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003435:Crotalid Venoms / Q000737:chemistry*; D004487:Edema / Q000139:chemically induced; D006801:Humans; D050830:L-Amino Acid Oxidase / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D051379:Mice; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010974:Platelet Aggregation / Q000187:drug effects; D013997:Time Factors
D000900:Anti-Bacterial Agents; D003435:Crotalid Venoms; D050830:L-Amino Acid Oxidase