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Research in Pharmaceutical Sciences، جلد ۲۰، شماره ۲، صفحات ۲۰۷-۲۱۷
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عنوان انگلیسی Isolation, characterization, and expression of the Montivipera raddei (Caucasus viper) venom metalloprotease-like protein
چکیده انگلیسی مقاله Background and purpose: Snake venom contains numerous proteolytic enzymes, including metalloproteases. This study aimed to isolate, characterize the metalloproteinase from Montivipera raddei venom and insert its cDNA into a host cell genome for expressing the protein's potential use as a coagulation factor. Experimental approach: Initially, M. raddei venom was analyzed using SDS-PAGE and ion-exchange chromatography. All purified fractions were assessed using a prothrombin time (PT) assay. Immunoblot analysis and MALDI-TOF/TOF MS spectrometry confirmed the metalloproteinase active domain in the obtained fraction. All of the confirmations were studied using the I-TASSER server. To produce the recombinant enzyme, metalloproteinase cDNA was isolated from the total RNA of the Caucasus viper venom gland tissue and cloned into pEX-A2-H plasmid, containing homologous sequences with CHO-S cell genome. Also, it was confirmed by western blotting and PT test. Findings/Results: Two fractions from M. raddei venom had PT values of 6 and 21 s, respectively. We confirmed the presence of a 25-KDa procoagulant metalloprotease (Met). The results indicated successful expression of the Met protein in the recombinant CHO-S cells. The Met protein was structurally similar to the target in the PDB ID. 2e3x, regulatory subunit of the blood coagulation factor X- and IX-activating enzyme. The Met protein also contained a similar binding domain with 60 amino acids, comparable to those in E. carinatus . and E. pyramidum leakeyi metalloproteinases. Conclusion and implications: The Met protein may be a potent candidate recombinant coagulant drug. Remarkably, the thrombin time for this protein was 3 s.    
کلیدواژه‌های انگلیسی مقاله Cloning,Chromatography,Metalloproteinase,Venom proteins.
نویسندگان مقاله | Zohreh Jafari
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, I.R. Iran.


| Behnam Sedaghati
Department of Environmental Science, Faculty of Natural Resources, University of Tehran, Tehran, I.R. Iran.


| Mohammad Kaboli
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, I.R. Iran.


| Bahram Kazemi
Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, I.R. Iran. Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, I.R. Iran.


| Mojgan Bandehpour


نشانی اینترنتی http://rps.mui.ac.ir/index.php/jrps/article/view/2301
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زبان مقاله منتشر شده en
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نوع مقاله منتشر شده Original Article
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