Academic Journal
Isolation and characterization of the first phosphodiesterase (Bj-PDE) from the venom of Bothrops jararacussu snake.
| العنوان: | Isolation and characterization of the first phosphodiesterase (Bj-PDE) from the venom of Bothrops jararacussu snake. |
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| المؤلفون: | Gobbi Amorim, Fernanda, Silva, Thiago Abrahão, de Oliveira Almeida, Gabriela, Redureau, Damien, Cabral, Hamilton, Quinton, Loïc, Sampaio, Suely Vilela |
| المصدر: | International Journal of Biological Macromolecules, 235, 123793 (2023-04-30) |
| بيانات النشر: | Elsevier B.V., 2023. |
| سنة النشر: | 2023 |
| مصطلحات موضوعية: | Bothrops jararacussu, Native toxin, Phosphodiesterase, Purification, Venom, Crotalid Venoms, Phosphoric Diester Hydrolases, Edetic Acid, Animals, Phosphoric Diester Hydrolases/chemistry, Chromatography, Crotalid Venoms/chemistry, Bothrops, Structural Biology, Biochemistry, Molecular Biology, General Medicine, Physical, chemical, mathematical & earth Sciences, Chemistry, Physique, chimie, mathématiques & sciences de la terre, Chimie |
| الوصف: | Phosphodiesterases are exonucleases that sequentially hydrolyse phosphodiester bonds of polynucleotides from the 3'-end and release 5-mononucleotides. After more than one decade without any advance in the study of Bothropic phosphodiesterases, we described here the isolation of the first phosphodiesterase from Bothrops jararacussu, which we named Bj-PDE. A five-step column chromatography procedure (size exclusion, hydrophobic interaction, cation exchange, lentil lectin affinity, and blue sepharose affinity) enabled isolation of Bj-PDE with preserved and stable enzymatic activity (bis(p-nitrophenyl) phosphate substrate), Km = 6.9 mM (± 0.7 mM), kcat/Km = 1.7 × 104 M-1 s-1 (± 0.2 × 104 M-1 s-1), MW = 116 kDa (SDS-PAGE), optimum activity around 45 °C at pH 8.0, and stability for 81 days at different storage temperatures (8, -20, and - 80 °C). Ca2+ and Mg2+ ions positively influenced Bj-PDE activity, while EDTA had the opposite action. Zn2+ restored >50 % of enzyme activity after its inhibition by EDTA. The Bj-PDE partial sequence identified by mass spectrometry was very similar to the sequence of BATXPDE1 from Bothrops atrox, which was evolutionarily close to this new PDE. Therefore, our study represents an important progress on the isolation of this minor toxin and sheds new lights on the properties and bioprospection of bothropic phosphodiesterases. |
| نوع الوثيقة: | journal article http://purl.org/coar/resource_type/c_6501 article peer reviewed |
| اللغة: | English |
| Relation: | https://api.elsevier.com/content/article/PII:S0141813023006876?httpAccept=text/xml; urn:issn:0141-8130; urn:issn:1879-0003 |
| DOI: | 10.1016/j.ijbiomac.2023.123793 |
| URL الوصول: | https://orbi.uliege.be/handle/2268/302094 |
| Rights: | restricted access http://purl.org/coar/access_right/c_16ec info:eu-repo/semantics/restrictedAccess |
| رقم الانضمام: | edsorb.302094 |
| قاعدة البيانات: | ORBi |
| DOI: | 10.1016/j.ijbiomac.2023.123793 |
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