人胎肺细胞的条件化培养液中两种类型纤溶酶原活化物的分离

Isolation of Two Types of Plasminogen Activator from Human Fetal Lung Cell Conditioned Medium

用正常人胎肺细胞体外培养，从其培养液中分离纤溶酶原活化物（ＰＡ），通过ＣＭ－ＳｅｐｈａｄｅｘＣ－５０层析，硫酸铵沉淀和Ｆｉｂｒｉｎ－Ｓｅｐｈａｒｏｓｅ，Ｌｙｓｉｎｅ－Ｓｅｐｈａｒｏｓｅ亲和层析及ＳｅｐｈａｄｅｘＧ－５０凝胶过滤等步骤，从１０．５ｌ条件培养液中分离纯化得到两种类型的纤溶酶原活化物，ｔ－ＰＡ９０μｇ，ｕ－ＰＡ８００μｇ．在还原条件下ＳＤＳ－ＰＡＧＥ均显示单带，分子量ｔ－ＰＡ为７２ｋＤ，ｕ－ＰＡ为５４ｋＤ，纤溶比活分别为１５６０００ＩＵ／ｍｇ蛋白和１０６０００ＩＵ／ｍｇ蛋白．

Two types of plasminogen activator(tissue type PA and urokinase type PA)are important regulators in fibrinolysis.A method was adopted to isolate and purify t PA from normal human fetal lung cell conditioned medium.Its procedures consisted of CM Sephadex C 50 chromatography,ammonium sulfate precipitation,Fibrin Sepharose,Lysine Sepharose affinity chromatography and gel filtration on Sephadex G 50.The final products of t PA 90 μg and u PA 800 μg from 10 5 L conditioned medium of human fetal lung cells had molecular weights of 72 kD and 54 kD respectively as estimated by SDS PAGE in the presence of the reducing reagent,and had specific activities of 156 000 IU/mg protein and 106 000 IU/mg protein respectively.