背景:来自蚯蚓的Lu激酶,已知龙具有纤溶酶,由于其溶解纤维蛋白的能力,其具有作为治疗药物的潜力。目前的研究旨在从L.rubellus中纯化Lumbrokase并鉴定其蛋白质成分。
方法:当地蚯蚓的水提取物揭示了几种蛋白质。因此,为了识别它的蛋白质成分,在鉴定之前通过HiPrepDEAE快速流动和蛋白质组学分析进行纯化。使用二维凝胶电泳(2DE)和电喷雾电离质谱分析的组合来鉴定纯化的级分。
结果:纯化的组分含有5条蛋白带,即F25-1、F25-2、F85-1、F85-2和F85-3,其显示出强的纤维蛋白原溶解活性。F25组分显示纤溶活性为974.85U/mg,而F85组分显示出更高的活性,为1,484.11U/mg。F85-1,F85-2和F85-3的分子量为42.6kDa,27.03kDa,和14kDa,分别被鉴定为Lumbrokinase同工酶。
结论:这项初步研究表明,F25和F85组分与已发表的纤溶蛋白酶-1和肺激酶相似,分别,就其氨基酸序列而言。
BACKGROUND: Lumbrokinase derived from earthworms, Lumbricus rubellus is known to have fibrinolytic enzymes that have potential as therapeutic drugs due to its ability to dissolve fibrin. The current study is aimed to purify the Lumbrokinase from L. rubellus and identify its protein component.
METHODS: Water extract of local earthworm Lumbricus rubellus revealed several proteins. Therefore, to identify its protein component, purification through HiPrep DEAE fast flow and proteomic analysis were conducted prior to identifications. A combination of two-dimension gel electrophoresis (2DE) and electrospray ionization mass spectrometry analysis was used to identify the purified fractions.
RESULTS: The purified fractions contain five protein bands, namely F25-1, F25-2, F85-1, F85-2, and F85-3, which displayed strong fibrinogenolytic activity. F25 fractions showed fibrinogenolytic activity of 974.85 U/mg, while F85 fractions showed higher activity of 1,484.11 U/mg. Fractions F85-1, F85-2, and F85-3 showed molecular weights of 42.6 kDa, 27.03 kDa, and 14 kDa, respectively and were identified as Lumbrokinase iso-enzymes.
CONCLUSIONS: This preliminary study indicates that the F25 and F85 fractions are similar to published fibrinolytic protease-1 and lumbrokinase, respectively, in terms of their amino acid sequence.