Characterization and amino-terminal sequence of phospholipase A2-II from the venom of Agkistrodon bilineatus (common cantil).
1. Phospholipase A2 was isolated from the venom of Agkistrodon bilineatus by Sephadex G-75 and CM-Cellulose column chromatographies. 2. The purified phospholipase A2 gave a single band on disc polyacrylamide gel electrophoresis, sodium dodecyl sulfate polyacrylamide gel electrophoresis and ODS-HPLC. 3. The enzyme preparation had a mol. wt of 14,000, isoelectric point of pH 10.12 and possessed 121 amino acid residues. 4. The enzyme hydrolyzed the phospholipids phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl inositol and phosphatidyl serine. 5. The contraction of mouse diaphragm was inhibited by phospholipase A2-II. 6. Phospholipase A2 activity of this preparation was inhibited by ethylenediamine tetraacetic acid, ethyleneglycol (beta-aminoethyl) N,N,N',N'-tetraacetic acid, p-bromophenacyl bromide or N-bromosuccinimide, but not by iodoacetic acid or diisopropyl fluorophosphate. 7. The amino-terminal sequence of the PLA2-II was determined.
Nikai
T
T
Department of Microbiology, Faculty of Pharmacy, Meijo University, Nagoya, Japan.
Komori
Y
Y
Ohara
A
A
Yagihashi
S
S
Ohizumi
Y
Y
Sugihara
H
H
D000595:Amino Acid Sequence; D000818:Animals; D003435:Crotalid Venoms / Q000201:enzymology*; D004795:Enzyme Stability; D054501:Group II Phospholipases A2; D006358:Hot Temperature; D007928:Lethal Dose 50; D051379:Mice; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000737:chemistry* / Q000302:isolation & purification / Q000633:toxicity; D054467:Phospholipases A2; D013379:Substrate Specificity
C031442:Agkistrodon venoms; D003435:Crotalid Venoms; D010741:Phospholipases A; D054501:Group II Phospholipases A2; D054467:Phospholipases A2