Volume 11, Issue 43 (7-2021)
NCMBJ 2021, 11(43): 39-45 |
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Mohammadi N, Bandehpour M, Sotoodehnejadnematalahi F, Kazemi B. Recombinant Production of metalloproteinase domain of Ecarin as the main activator of prothrombin. NCMBJ 2021; 11 (43) :39-45
URL: http://ncmbjpiau.ir/article-1-1393-en.html
URL: http://ncmbjpiau.ir/article-1-1393-en.html
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
Abstract: (2531 Views)
Aim and Background: Today, many patients with coagulation disorders due to defective or non-functioning coagulation factors and the use of various blood thinners need to check their prothrombin total and quantify determination of direct thrombin inhibitors. Despite the study of the ecarin from Echis carinatus venom as a direct activator of prothrombin, there have been not investigations into the active site of this enzyme.
Material and methods: Ecarin metalloproteinase domain (639 bp) was synthesized into the pcDNA3.1. Recombinant plasmid was transfected into HEK293 cell line. Protein expression was evaluated using SDS-PAGE and its function in reaction with prothrombin and using a prothrombin time test was measured.
Results: The results showed that the metalloproteinase domain of ecarin produced in the HEK293, like whole ecarin, was able to activate prothrombin to thrombin; but its activity was slow than one produced in the prokaryotic system.
Conclusion: In this study, the recombinant ecarin metalloproteinase produced in HEK293 cells, like whole ecarin, was able to activate prothrombin to thrombin and was introduced for the first time as a suitable alternative to both natural and recombinant ecarin in methods based on prothrombin to thrombin conversion.
Material and methods: Ecarin metalloproteinase domain (639 bp) was synthesized into the pcDNA3.1. Recombinant plasmid was transfected into HEK293 cell line. Protein expression was evaluated using SDS-PAGE and its function in reaction with prothrombin and using a prothrombin time test was measured.
Results: The results showed that the metalloproteinase domain of ecarin produced in the HEK293, like whole ecarin, was able to activate prothrombin to thrombin; but its activity was slow than one produced in the prokaryotic system.
Conclusion: In this study, the recombinant ecarin metalloproteinase produced in HEK293 cells, like whole ecarin, was able to activate prothrombin to thrombin and was introduced for the first time as a suitable alternative to both natural and recombinant ecarin in methods based on prothrombin to thrombin conversion.
Type of Study: Research Article |
Subject:
Cellular and molecular
Received: 2021/02/13 | Accepted: 2021/06/20 | Published: 2021/07/1
Received: 2021/02/13 | Accepted: 2021/06/20 | Published: 2021/07/1
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