دورية أكاديمية
Structural and Functional Characterization of New SsoPox Variant Points to the Dimer Interface as a Driver for the Increase in Promiscuous Paraoxonase Activity
العنوان: | Structural and Functional Characterization of New SsoPox Variant Points to the Dimer Interface as a Driver for the Increase in Promiscuous Paraoxonase Activity |
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المؤلفون: | Yoko Suzumoto, Orly Dym, Giovanni N. Roviello, Franz Worek, Joel L. Sussman, Giuseppe Manco |
المصدر: | International Journal of Molecular Sciences, Vol 21, Iss 5, p 1683 (2020) |
بيانات النشر: | MDPI AG, 2020. |
سنة النشر: | 2020 |
المجموعة: | LCC:Biology (General) LCC:Chemistry |
مصطلحات موضوعية: | molecular evolution, phosphotriesterase-like-lactonases (plls), organophosphate (op) compounds, nerve agents, enzymatic detoxification/remediation, Biology (General), QH301-705.5, Chemistry, QD1-999 |
الوصف: | Increasing attention is more and more directed toward the thermostable Phosphotriesterase-Like-Lactonase (PLL) family of enzymes, for the efficient and reliable decontamination of toxic nerve agents. In the present study, the DNA Staggered Extension Process (StEP) technique was utilized to obtain new variants of PLL enzymes. Divergent homologous genes encoding PLL enzymes were utilized as templates for gene recombination and yielded a new variant of SsoPox from Saccharolobus solfataricus. The new mutant, V82L/C258L/I261F/W263A (4Mut) exhibited catalytic efficiency of 1.6 × 105 M−1 s−1 against paraoxon hydrolysis at 70°C, which is more than 3.5-fold and 42-fold improved in comparison with C258L/I261F/W263A (3Mut) and wild type SsoPox, respectively. 4Mut was also tested with chemical warfare nerve agents including tabun, sarin, soman, cyclosarin and VX. In particular, 4Mut showed about 10-fold enhancement in the hydrolysis of tabun and soman with respect to 3Mut. The crystal structure of 4Mut has been solved at the resolution of 2.8 Å. We propose that, reorganization of dimer conformation that led to increased central groove volume and dimer flexibility could be the major determinant for the improvement in hydrolytic activity in the 4Mut. |
نوع الوثيقة: | article |
وصف الملف: | electronic resource |
اللغة: | English |
تدمد: | 1422-0067 |
Relation: | https://www.mdpi.com/1422-0067/21/5/1683; https://doaj.org/toc/1422-0067 |
DOI: | 10.3390/ijms21051683 |
URL الوصول: | https://doaj.org/article/37922a3a3412494c88b970a78f470696 |
رقم الأكسشن: | edsdoj.37922a3a3412494c88b970a78f470696 |
قاعدة البيانات: | Directory of Open Access Journals |
تدمد: | 14220067 |
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DOI: | 10.3390/ijms21051683 |