一种新型蚯蚓纤溶酶组分的部分性质研究  

Characterization of a new fibrinolytic enzyme from Eisenia foelida

作　　者：李红[1] 隋洪艳[1] 杨立清[1] 严宜明[1] 石男[1] 

机构地区：[1]北京生物制品研究所免疫诊断研究室,北京100024

出　　处：《中国生化药物杂志》2004年第6期328-330,共3页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的获得一种高效的溶栓药物。方法从赤子爱胜蚓 (Eiseniafoelida)中共提取 6种纤溶酶组分 ,分别命名为F1～F6。取F1用Lowry法测定蛋白质浓度 ,SDS PAGE鉴定纯度及表观相对分子质量 (Mr) ,纤维蛋白平板法测定其纤溶活性 ,测定其水解BAEE(Nα 苯甲酰 L 精氨酸乙酯盐酸盐 )、血浆纤溶酶特异性底物ChromozymPL(苄氧羰酰甘氨酰脯氨酰精氨酰对硝基苯胺盐酸盐 )及组织型纤溶酶原激活剂Chromozymt PA(N 甲磺酰苯丙酰甘氨酰精氨酰对硝基苯胺盐酸盐 )的活性 ,并进行N端氨基酸序列测定。结果F1纯度为 10 0 % ,表观Mr 为 2 85 0 0 ,总纤溶活性为 6 5 .5 1× 10 3 mm2 /mg ,水解BAEE的米氏常数 (Km)为 2 .80 87× 10 -2 mol/L。对chromozymPL及chromozymt PA的亲和力较低。F1的N端氨基酸序列为IIGGSNASPGEFPWQ ,且对天然凝血块有较强的溶解作用。结论F1为纯度很高的单一组分 ,纤溶活性较高。PurposeTo obtain an effective drug for thrombolysis.MethodsSix components were isolated and purified from earthworm Eisenia foelida and named F1…F6 . The protein concentration of F1 was determined by Lowry method. The purity and M r of F1 were estimated by SDS-poly acrylamide gel deectrophoresis(SDS-PAGE). The fibrinolytic activity was determined by Fibrin plate method. BAEE, Chromozym PL and Chromozym t-PA were used as specific substrate to obtain the ΔA/(min.g) and Km of F1 .The sequence of N-terminus 15 amino acids was determined by Applied Biosystem 491 Protein Sequencer.ResultsThe purity and M r of F1 were 100% and 28 500 respectively. F1 displayed a strong fibrinolytic activity with fibrin plate method but could weakly hydrolyze Chromozym PL and Chromozym t-PA.The Km of Chromozym PL and Chromozym t-PA could'nt be detected .The Km of BAEE decomposed by F1 was 2.808 7×10 -2 mol/L. Its N-terminal amino acid sequence was IIGGSNASPGEFPWQ.ConclusionA new and effective fibrinolytic enzyme is obtained.

关 键 词：赤子爱胜蚓 纤溶活性 米氏常数 序列测定 

分 类 号：Q556[生物学—生物化学] R392-33[医药卫生—免疫学]