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

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. METHOD...

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Autores principales: Stephani, Laurentia, Rahayu, Puji, Retnoningrum, Debbie, Suhartono, Maggy Thenawidjaja, Rachmawati, Heni, Tjandrawinata, Raymond R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10165752/
https://www.ncbi.nlm.nih.gov/pubmed/37158880
http://dx.doi.org/10.1186/s12953-023-00206-9
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author Stephani, Laurentia
Rahayu, Puji
Retnoningrum, Debbie
Suhartono, Maggy Thenawidjaja
Rachmawati, Heni
Tjandrawinata, Raymond R.
author_facet Stephani, Laurentia
Rahayu, Puji
Retnoningrum, Debbie
Suhartono, Maggy Thenawidjaja
Rachmawati, Heni
Tjandrawinata, Raymond R.
author_sort Stephani, Laurentia
collection PubMed
description 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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spelling pubmed-101657522023-05-09 Purification and proteomic analysis of potent fibrinolytic enzymes extracted from Lumbricus rubellus Stephani, Laurentia Rahayu, Puji Retnoningrum, Debbie Suhartono, Maggy Thenawidjaja Rachmawati, Heni Tjandrawinata, Raymond R. Proteome Sci Research 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. BioMed Central 2023-05-08 /pmc/articles/PMC10165752/ /pubmed/37158880 http://dx.doi.org/10.1186/s12953-023-00206-9 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Stephani, Laurentia
Rahayu, Puji
Retnoningrum, Debbie
Suhartono, Maggy Thenawidjaja
Rachmawati, Heni
Tjandrawinata, Raymond R.
Purification and proteomic analysis of potent fibrinolytic enzymes extracted from Lumbricus rubellus
title Purification and proteomic analysis of potent fibrinolytic enzymes extracted from Lumbricus rubellus
title_full Purification and proteomic analysis of potent fibrinolytic enzymes extracted from Lumbricus rubellus
title_fullStr Purification and proteomic analysis of potent fibrinolytic enzymes extracted from Lumbricus rubellus
title_full_unstemmed Purification and proteomic analysis of potent fibrinolytic enzymes extracted from Lumbricus rubellus
title_short Purification and proteomic analysis of potent fibrinolytic enzymes extracted from Lumbricus rubellus
title_sort purification and proteomic analysis of potent fibrinolytic enzymes extracted from lumbricus rubellus
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10165752/
https://www.ncbi.nlm.nih.gov/pubmed/37158880
http://dx.doi.org/10.1186/s12953-023-00206-9
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