Kumada T, Abiko Y
Thromb Res. 1984 Oct 15;36(2):143-52. doi: 10.1016/0049-3848(84)90336-0.
alpha 2-Plasmin inhibitor was purified from rat plasma by the procedures including sequential lysine-Sepharose chromatography, ammonium sulfate precipitation and plasminogen-Sepharose, DEAE-Sephadex, concanavalin A-Sepharose, and Sephadex G-200 chromatographies. The final preparation was homogeneous as judged by sodium dodecyl sulfate polyacrylamide gel- and immuno-electrophoresis. It was a single polypeptide chain with a molecular weight of about 7.1 X 10(4) daltons and was found to possess fast-acting antiplasmin activity. There existed other molecular forms of alpha 2-plasmin inhibitor-related antigen with molecular weight of 6.3 X 10(4) and 6.1 X 10(4) daltons. The latter was found to be inactive in binding to plasminogen and in inhibiting plasmin activity. Normal plasma level of alpha 2-plasmin inhibitor in rats was immunologically determined to be 7.9 +/- 0.8 mg/dl. Indirect evidence was obtained that rat alpha 2-plasmin inhibitor was incorporated into cross-linked fibrin during blood clotting.
通过包括赖氨酸-琼脂糖凝胶柱层析、硫酸铵沉淀以及纤溶酶原-琼脂糖凝胶柱层析、DEAE-葡聚糖凝胶柱层析、伴刀豆球蛋白A-琼脂糖凝胶柱层析和葡聚糖凝胶G-200柱层析等步骤，从大鼠血浆中纯化出α2-纤溶酶抑制剂。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳和免疫电泳判断，最终制品为均一的。它是一条单多肽链，分子量约为7.1×10⁴道尔顿，具有快速起效的抗纤溶酶活性。还存在分子量为6.3×10⁴和6.1×10⁴道尔顿的其他分子形式的α2-纤溶酶抑制剂相关抗原。发现后者在与纤溶酶原结合及抑制纤溶酶活性方面无活性。通过免疫测定法确定，大鼠血浆中α2-纤溶酶抑制剂的正常水平为7.9±0.8mg/dl。获得了间接证据，表明大鼠α2-纤溶酶抑制剂在血液凝固过程中被整合到交联纤维蛋白中。
