دورية أكاديمية
Structure of a diguanylate cyclase from Thermotoga maritima: insights into activation, feedback inhibition and thermostability.
العنوان: | Structure of a diguanylate cyclase from Thermotoga maritima: insights into activation, feedback inhibition and thermostability. |
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المؤلفون: | Angeline Deepthi, Chong Wai Liew, Zhao-Xun Liang, Kunchithapadam Swaminathan, Julien Lescar |
المصدر: | PLoS ONE, Vol 9, Iss 10, p e110912 (2014) |
بيانات النشر: | Public Library of Science (PLoS), 2014. |
سنة النشر: | 2014 |
المجموعة: | LCC:Medicine LCC:Science |
مصطلحات موضوعية: | Medicine, Science |
الوصف: | Large-scale production of bis-3'-5'-cyclic-di-GMP (c-di-GMP) would facilitate biological studies of numerous bacterial signaling pathways and phenotypes controlled by this second messenger molecule, such as virulence and biofilm formation. C-di-GMP constitutes also a potentially interesting molecule as a vaccine adjuvant. Even though chemical synthesis of c-di-GMP can be done, the yields are incompatible with mass-production. tDGC, a stand-alone diguanylate cyclase (DGC or GGDEF domain) from Thermotoga maritima, enables the robust enzymatic production of large quantities of c-di-GMP. To understand the structural correlates of tDGC thermostability, its catalytic mechanism and feedback inhibition, we determined structures of an active-like dimeric conformation with both active (A) sites facing each other and of an inactive dimeric conformation, locked by c-di-GMP bound at the inhibitory (I) site. We also report the structure of a single mutant of tDGC, with the R158A mutation at the I-site, abolishing product inhibition and unproductive dimerization. A comparison with structurally characterized DGC homologues from mesophiles reveals the presence of a higher number of salt bridges in the hyperthermophile enzyme tDGC. Denaturation experiments of mutants disrupting in turn each of the salt bridges unique to tDGC identified three salt-bridges critical to confer thermostability. |
نوع الوثيقة: | article |
وصف الملف: | electronic resource |
اللغة: | English |
تدمد: | 1932-6203 |
Relation: | http://europepmc.org/articles/PMC4215984?pdf=render; https://doaj.org/toc/1932-6203 |
DOI: | 10.1371/journal.pone.0110912 |
URL الوصول: | https://doaj.org/article/cd734726e32e44b097dde757b0f06959 |
رقم الأكسشن: | edsdoj.734726e32e44b097dde757b0f06959 |
قاعدة البيانات: | Directory of Open Access Journals |
تدمد: | 19326203 |
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DOI: | 10.1371/journal.pone.0110912 |