Analysis of fibrinolytic activation mechanism by staphylokinase in gene deficient mice

葡萄激酶（SAK）通过与纤溶酶原（猪）或纤溶酶形成复合物来表达纤溶酶原激活物（PA）的活性。虽然这种PA活性被alpha2-antiplasmin （alpha2-AP）抑制，但在纤维蛋白存在时，其活性不受alpha2-AP的抑制。利用SAKs突变体、合成的SAK肽、几种动物的纯化猪和基因缺陷小鼠的血浆研究了SAK与猪的相互作用。突变SAKs与猪的相互作用。SAK nh2末端氨基酸区缺失突变体由于缺失前14、18和25个氨基酸区（14、18和25）而收缩。SAK的11-14氨基酸区与glu -plg结合后表达PA活性是必需的(2)。合成SAK肽与猪的相互作用。合成的SAK肽在SAK氨基酸序列中包含10-30个氨基酸。合成的SAK肽不显示PA活性。然而，合成的SAK非十肽（Glu22-Leu4O）与Glupig猪结合，增强了猪对天然SAK的激活。α 2- ap和猪基因缺陷小鼠。通过克隆和靶向小鼠α 2- ap基因获得α 2- ap基因缺陷小鼠（α 2- ap -/-）。猪基因缺陷小鼠（pig -/-）建立了猪基因在小鼠体内的繁育方法和测定方法。SAK与纯化猪缓冲液、α 2- ap或猪缺陷小鼠血浆的相互作用。猪从人、鼠、兔或牛的血浆中纯化。在缓冲液和α 2- ap -/-或猪-/-小鼠血浆中研究了SAK与纯化猪之间的相互作用。这些结果表明：(1)小鼠或牛plg不与SAK发生反应，(2)SAK-人piasmin复合物激活小鼠或牛plg，(3)小鼠plg的激活被小鼠α 2- ap抑制。

Staphylokinase (SAK) expresses plasminogen activator (PA) activity by forming a complex with plasminogen (Pig) or plasmin. Although this PA activity is inhibited by alpha2-antiplasmin (alpha2-AP), its activity is protected against the inhibition by alpha2-AP in the presnce of fibrin. The interaction between SAK and Pig were investigated using mutant SAKs, synthetic SAK peptides, purified pigs of several animals and gene deficient mice plasma.1). The interaction between mutant SAKs and pig.The deleted mutants of NH2-terminal amino acid region of SAK contracted by lacking the first 14, 18 and 25 amino acid region ( 14 SAK, 18 SAK and 25 SAK). The11-14 amino acid region of SAK was necessary to express PA activity after binding with Glu-plg.2). The interaction between synthetic SAK peptides and pig.The SAK peptides synthesized peptides consisted 10-30 amino acid in SAK amino acid sequence. The synthetic SAK peptides did not show PA activity. However, the synthetic nonadecapeptide (Glu22-Leu4O) of SAK bound to Glupig pig and enhanced pig activation by native SAK.3). The alpha2-AP and Pig gene deficient mice.The alpha2-AP gene deficient mice (alpha2-AP-/-) obtained by cloning and targeting of murine alpha2-AP gene. The pig gene deficient mice (Pig-/-)established breeding method and assay methods of pig gene in mice.4). The interaction between SAK and purified pigs in buffer and in alpha2-AP or Pig deficient mice plasma.The pigs purified from plasma of human, mice, rabbit or bovine. The interaction between SAK and purified pigs investigated in buffer and in alpha2-AP-/- or Pig-/- mice plasma. These findings indicate that (1) murine or bovine plg dose not react with SAK, (2) SAK-human piasmin complex activates murine or bovine plg, and (3) this murine plg activation is inhibited by murine alpha2-AP.