Academic Journal
A Single Point Mutation Converts GH84 O-GlcNAc Hydrolases into Phosphorylases: Experimental and Theoretical Evidence
| العنوان: | A Single Point Mutation Converts GH84 O-GlcNAc Hydrolases into Phosphorylases: Experimental and Theoretical Evidence |
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| المؤلفون: | Teze, David, Coines, Joan, Raich, Lluís, Kalichuk, Valentina, Solleux, Claude, Tellier, Charles, André-Miral, Corinne, Svensson, Birte, Rovira, Carme |
| المصدر: | Teze , D , Coines , J , Raich , L , Kalichuk , V , Solleux , C , Tellier , C , André-Miral , C , Svensson , B & Rovira , C 2020 , ' A Single Point Mutation Converts GH84 O-GlcNAc Hydrolases into Phosphorylases: Experimental and Theoretical Evidence ' , Journal of the American Chemical Society , vol. 142 , no. 5 , pp. 2120-2124 . https://doi.org/10.1021/jacs.9b09655 |
| سنة النشر: | 2020 |
| المجموعة: | Technical University of Denmark: DTU Orbit / Danmarks Tekniske Universitet |
| الوصف: | Glycoside hydrolases and phosphorylases are two major classes of enzymes responsible for the cleavage of glycosidic bonds. Here we show that two GH84 O-GlcNAcase enzymes can be converted to efficient phosphorylases by a single point mutation. Noteworthy, the mutated enzymes are over 10-fold more active than naturally occurring glucosaminide phosphorylases. We rationalize this novel transformation using molecular dynamics and QM/MM metadynamics methods, showing that the mutation changes the electrostatic potential at the active site and reduces the energy barrier for phosphorolysis by 10 kcal·mol -1 . In addition, the simulations unambiguously reveal the nature of the intermediate as a glucose oxazolinium ion, clarifying the debate on the nature of such a reaction intermediate in glycoside hydrolases operating via substrate-assisted catalysis. |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| Relation: | https://orbit.dtu.dk/en/publications/6a54425d-c973-4232-af8a-1917a42e730c |
| DOI: | 10.1021/jacs.9b09655 |
| الاتاحة: | https://orbit.dtu.dk/en/publications/6a54425d-c973-4232-af8a-1917a42e730c https://doi.org/10.1021/jacs.9b09655 https://backend.orbit.dtu.dk/ws/files/205496678/bsog_jacs.9b09655.pdf |
| Rights: | info:eu-repo/semantics/openAccess |
| رقم الانضمام: | edsbas.CFDDB673 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1021/jacs.9b09655 |
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