A fibrinolytic protease AfeE from Streptomyces sp. CC5, with potent thrombolytic activity in a mouse model.
A fibrinolytic protease AfeE from Streptomyces sp. CC5, with potent thrombolytic activity in a mouse model.
复制标题
DOI：
10.1016/j.ijbiomac.2015.12.059
复制
发表时间：
2016-04
影响因子：
8.2
通讯作者：
Zhibin Sun;Pingping Liu;Guangyan Cheng;Biying Zhang;W. Dong;Xing-li Su;Y. Huang;Z. Cui;Yi Ko
中科院分区：
文献类型：
--
作者：
Zhibin Sun;Pingping Liu;Guangyan Cheng;Biying Zhang;W. Dong;Xing-li Su;Y. Huang;Z. Cui;Yi Ko
纤溶蛋白酶在心血管疾病治疗中具有潜在应用。从链霉菌中纯化出一种具有强溶栓活性的新型纤溶蛋白酶AfeE。 CC5。 AfeE 在 40°C、pH 值 7.0–12.0 范围内表现出最大活性。丝氨酸蛋白酶抑制剂苯甲磺酰氟、大豆胰蛋白酶抑制剂、甲苯磺酰-L-赖氨酸氯甲基酮和甲苯磺酰-L-苯丙氨酸氯甲基酮对其有强烈抑制作用。 Cu2+、Co2+和Zn2+部分抑制该酶的活性。 AfeE 对纤维蛋白表现出比纤维蛋白原更高的底物特异性，这在纤溶酶中很少有报道。 AfeE 在角叉菜胶诱导的小鼠尾部血栓形成模型中也表现出高溶栓活性。 AfeE 延长大鼠血液中的凝血酶原时间、活化部分凝血活酶时间和凝血酶时间。出血时间测定表明，1 毫克/千克剂量的 AfeE 不会延长小鼠的出血时间。在人脐静脉内皮细胞中，320 μg/孔的 AfeE 未观察到急性细胞毒性。 TheafeE基因是从Streptomycesp的基因组中克隆的。 CC5。高分辨率质谱分析显示，全长 AFE-CC5E 含有 434 个氨基酸，通过翻译后修饰加工成包含 284 个氨基酸的成熟形式。这些结果表明 AfeE 是抗血栓药物开发的潜在候选者。
Fibrinolytic proteases have potential applications in cardiovascular disease therapy. A novel fibrinolytic protease, AfeE, with strong thrombolytic activity was purified fromStreptomycessp. CC5. AfeE displayed maximum activity at 40 °C in the pH range of 7.0–12.0. It was strongly inhibited by serine protease inhibitor phenylmethanesulfonylfluoride, soybean trypsin inhibitor, tosyl-l-lysine chloromethyl ketone and tosyl-l-phenylalanine chloromethyl ketone. The activity of the enzyme was partially inhibited by Cu2+, Co2+and Zn2+. AfeE exhibited higher substrate specificity for fibrin than fibrinogen, which has rarely been reported in fibrinolytic enzymes. AfeE also showed high thrombolytic activity in a carrageenan-induced mouse tail thrombosis model. AfeE prolonged prothrombin time, activated partial thromboplastin time, and thrombin time in rat blood. A bleeding time assay revealed that AfeE did not prolong bleeding time in mice at a dose of 1 mg/kg. No acute cytotoxicity was observed for AfeE at 320 μg/well in human umbilical vein endothelial cells. TheafeEgene was cloned from the genome ofStreptomycessp. CC5. Full-length AFE-CC5E contained 434 amino acids and was processed into a mature form consisting 284 amino acids by posttranslational modification, as revealed by high-resolution mass spectrometry analysis. These results indicate that AfeE is a prospective candidate for antithrombotic drug development.