双带腹链蛇（普通坎蒂尔）毒液中磷脂酶A2-II的特性及氨基末端序列

Characterization and amino-terminal sequence of phospholipase A2-II from the venom of Agkistrodon bilineatus (common cantil).

Nikai T, Komori Y, Ohara A, Yagihashi S, Ohizumi Y, Sugihara H

Department of Microbiology, Faculty of Pharmacy, Meijo University, Nagoya, Japan.

Int J Biochem. 1994 Jan;26(1):43-8. doi: 10.1016/0020-711x(94)90193-7.

Abstract

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.

摘要

通过葡聚糖凝胶G - 75和CM - 纤维素柱色谱法从双斑锦蛇毒液中分离出磷脂酶A2。2. 纯化后的磷脂酶A2在圆盘聚丙烯酰胺凝胶电泳、十二烷基硫酸钠聚丙烯酰胺凝胶电泳和ODS - HPLC上呈现单一谱带。3. 该酶制剂的分子量为14,000，等电点为pH 10.12，含有121个氨基酸残基。4. 该酶可水解磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰肌醇和磷脂酰丝氨酸。5. 磷脂酶A2 - II可抑制小鼠膈肌收缩。6. 该制剂的磷脂酶A2活性受到乙二胺四乙酸、乙二醇（β - 氨基乙基）N,N,N',N'-四乙酸、对溴苯甲酰溴或N - 溴代琥珀酰亚胺的抑制，但不受碘乙酸或二异丙基氟磷酸的抑制。7. 测定了PLA2 - II的氨基末端序列。