دورية أكاديمية
Purification, Characterization and Evaluation of the Anticoagulant Effect of an Uncompetitive Trypsin Inhibitor obtained from Bauhinia pulchella (Benth) Seeds.
| العنوان: | Purification, Characterization and Evaluation of the Anticoagulant Effect of an Uncompetitive Trypsin Inhibitor obtained from Bauhinia pulchella (Benth) Seeds. |
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| المؤلفون: | Roma RR; Department of Biochemistry and Molecular Biology, Federal University of Ceará, Fortaleza, CE, Brazil., Dias LP; Paulista School of Medicine, Support Foundation for the Federal University of São Paulo, Sao Paulo, Brazil., Santos ALE; Agrarian and Biodiversity Sciences Center, Federal University of Cariri, Crato, Ceará, Brazil., Silva RRS; Department of Biochemistry and Molecular Biology, Federal University of Ceará, Fortaleza, CE, Brazil., Santos MHC; Department of Biochemistry and Molecular Biology, Federal University of Ceará, Fortaleza, CE, Brazil., Rocha BAM; Department of Biochemistry and Molecular Biology, Federal University of Ceará, Fortaleza, CE, Brazil., Carneiro RF; Department of Fisheries Engineering, Universidade Federal do Ceará, Fortaleza, CE, Brazil., Nagano CS; Department of Fisheries Engineering, Universidade Federal do Ceará, Fortaleza, CE, Brazil., Sampaio AH; Department of Fisheries Engineering, Universidade Federal do Ceará, Fortaleza, CE, Brazil., Oliva MLV; Department of Biochemistry, Federal University of São Paulo, São Paulo, SP, Brazil., Silva CGL; Medical School, Federal University of Cariri, Barbalha, Ceará, Brazil., Souza ROS; Medical School, Federal University of Cariri, Barbalha, Ceará, Brazil., Teixeira CS; Agrarian and Biodiversity Sciences Center, Federal University of Cariri, Crato, Ceará, Brazil. |
| المصدر: | Current protein & peptide science [Curr Protein Pept Sci] 2024; Vol. 25 (2), pp. 172-182. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Bentham Science Publishers Country of Publication: United Arab Emirates NLM ID: 100960529 Publication Model: Print Cited Medium: Internet ISSN: 1875-5550 (Electronic) Linking ISSN: 13892037 NLM ISO Abbreviation: Curr Protein Pept Sci Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Saif Zone, Sharjah, U.A.E. : Bentham Science Publishers Original Publication: Hilversum, Netherlands ; Boca Raton, FL : Bentham Science Publishers, c2000- |
| مواضيع طبية MeSH: | Trypsin Inhibitors*/pharmacology , Trypsin Inhibitors*/chemistry , Bauhinia*/metabolism, Animals ; Cattle ; Trypsin/analysis ; Trypsin/chemistry ; Trypsin/metabolism ; Tandem Mass Spectrometry ; Seeds/chemistry ; Anticoagulants/pharmacology ; Anticoagulants/analysis ; Anticoagulants/chemistry |
| مستخلص: | Introduction: Trypsin inhibitors (TIs) have the ability to competitively or non-competitively bind to trypsin and inhibit its action. These inhibitors are commonly found in plants and are used in protease inhibition studies involved in biochemical pathways of pharmacological interest. Objectives: This work aimed to purify a trypsin inhibitor from Bauhinia pulchella seeds ( Bpu TI), describing its kinetic mechanism and anticoagulant effect. Methods: Affinity chromatography, protein assay, and SDS-PAGE were used to purify the inhibitor. Mass spectrometry, inhibition assays, and enzyme kinetics were used to characterize the inhibitor. In vitro assays were performed to verify its ability to prolong blood clotting time. Results: Affinity chromatography on a Trypsin-Sepharose 4B column gave a yield of 43.1. Bpu TI has an apparent molecular mass of 20 kDa with glycosylation (1.15%). Protein identification was determined by MS/MS, and Bpu TI showed similarity to several Kunitz-type trypsin inhibitors. Bpu TI inhibited bovine trypsin as an uncompetitive inhibitor with IC50 (3 x 10 -6 M) and Ki (1.05 x 10 -6 M). Additionally, Bpu TI showed high stability to temperature and pH variations, maintaining its activity up to 100ºC and in extreme pH ranges. However, the inhibitor was susceptible to reducing agents, such as DTT, which completely abolished its activity. Bpu TI showed an anticoagulant effect in vitro at a concentration of 33 μM, prolonging clotting time by 2.6 times. Conclusion: Our results suggest that Bpu TI can be a biological tool to be used in blood clotting studies. (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.) |
| فهرسة مساهمة: | Keywords: Antithrombotic; Bauhinia; blood coagulation; fabaceae; protein; seeds. |
| المشرفين على المادة: | 0 (Trypsin Inhibitors) EC 3.4.21.4 (Trypsin) 0 (Anticoagulants) |
| تواريخ الأحداث: | Date Created: 20230911 Date Completed: 20240129 Latest Revision: 20240129 |
| رمز التحديث: | 20240129 |
| DOI: | 10.2174/1389203724666230908114115 |
| PMID: | 37694793 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1875-5550 |
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| DOI: | 10.2174/1389203724666230908114115 |