Purification and proteomic analysis of potent fibrinolytic enzymes extracted from Lumbricus rubellus.

IF 2.1 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS Proteome Science Pub Date : 2023-05-08 DOI:10.1186/s12953-023-00206-9

Laurentia Stephani, Puji Rahayu, Debbie Retnoningrum, Maggy Thenawidjaja Suhartono, Heni Rachmawati, Raymond R Tjandrawinata

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引用次数: 1

Abstract

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.

Conclusion: 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.

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风疹蚓强效纤溶酶的纯化及蛋白质组学分析。

背景:蚓激酶来源于蚯蚓，风疹蚓具有纤维蛋白溶解酶，由于其溶解纤维蛋白的能力，具有作为治疗药物的潜力。本研究旨在从风疹乳杆菌中纯化蚓激酶并鉴定其蛋白组分。方法:从当地蚯蚓rubellus水提液中提取多种蛋白质。因此，为了鉴定其蛋白质成分，在鉴定前通过HiPrep DEAE快速流纯化并进行蛋白质组学分析。采用二维凝胶电泳(2DE)和电喷雾电离质谱分析相结合的方法对纯化组分进行鉴定。结果:纯化后的部分含有F25-1、F25-2、F85-1、F85-2、F85-3 5条蛋白带，具有较强的纤维蛋白原溶解活性。F25组纤维蛋白原溶解活性为974.85 U/mg, F85组活性更高，为1484.11 U/mg。F85-1、F85-2和F85-3的分子量分别为42.6 kDa、27.03 kDa和14 kDa，鉴定为蚓激酶同工酶。结论:本初步研究表明，F25和F85组分的氨基酸序列分别与已发表的纤维蛋白溶解蛋白酶-1和蚓激酶相似。

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来源期刊

Proteome Science 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

审稿时长

4.5 months

期刊介绍： Proteome Science is an open access journal publishing research in the area of systems studies. Proteome Science considers manuscripts based on all aspects of functional and structural proteomics, genomics, metabolomics, systems analysis and metabiome analysis. It encourages the submissions of studies that use large-scale or systems analysis of biomolecules in a cellular, organismal and/or environmental context. Studies that describe novel biological or clinical insights as well as methods-focused studies that describe novel methods for the large-scale study of any and all biomolecules in cells and tissues, such as mass spectrometry, protein and nucleic acid microarrays, genomics, next-generation sequencing and computational algorithms and methods are all within the scope of Proteome Science, as are electron topography, structural methods, proteogenomics, chemical proteomics, stem cell proteomics, organelle proteomics, plant and microbial proteomics. In spite of its name, Proteome Science considers all aspects of large-scale and systems studies because ultimately any mechanism that results in genomic and metabolomic changes will affect or be affected by the proteome. To reflect this intrinsic relationship of biological systems, Proteome Science will consider all such articles.