[Purification and characterization of L-amino acid oxidase from Agkistrodon halys pallas venom].
L-amino acid oxidase (LAO, EC 1.4.3.2) is widely found in snake venoms and is thought to contribute to the toxicity in envenoming. By using of Sephadex G-150, DEAE-Sepharose CL-6B and FPLC Superose 12 chromatography, a protein with L-amino acid oxidase activity was purified and characterized from Agkistrodon haly Pallas venom. Its molecular mass was 57 kD as determined by SDS-PAGE analysis under both reducing and non-reducing conditions, and its pI was about 4.9. The protein catalysed the stereospecific oxidative deamination of L-amino acid substrate. It inhibited the platelet aggregation induced by ADP and collagen dose-dependently, even at low concentrations of 0.2 micromol/L and 0.08 micromol/L, respectively. The LAO had antibacterial effect to E.coli K12D31, and the effective concentration was as low as 0.03 g/L. Furthermore, the LAO showed cytotoxicity in crystal violet assay and apoptosis-inducing activity in the A549 cells. After 24h treatment with 5 mg/L LAO, the typical DNA fragmentation pattern of apoptotic cells was observed by using of agrose gel electrophoresis.
Liu
J W
JW
Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031, China.
Chai
M Q
MQ
Du
X Y
XY
Song
J G
JG
Zhou
Y C
YC
D017836:Agkistrodon / Q000378:metabolism*; D000594:Amino Acid Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D003435:Crotalid Venoms / Q000201:enzymology*; D053938:DNA Fragmentation / Q000187:drug effects; D006801:Humans; D050830:L-Amino Acid Oxidase; D008970:Molecular Weight; D010974:Platelet Aggregation / Q000187:drug effects
D003435:Crotalid Venoms; D000594:Amino Acid Oxidoreductases; D050830:L-Amino Acid Oxidase