|Title||Thermodynamic Stability of the Transcription Regulator PaaR2 from Escherichia coli O157:H7.|
|Publication Type||Journal Article|
|Year of Publication||2019|
|Authors||De Bruyn, P., S. Hadži, A. Vandervelde, A. Konijnenberg, M. Prolič-Kalinšek, Y. G. - J. Sterckx, F. Sobott, J. Lah, L. Van Melderen, and R. Loris|
|Date Published||2019 Apr 23|
PaaR2 is a putative transcription regulator encoded by a three-component parDE-like toxin-antitoxin module from Escherichia coli O157:H7. Although this module's toxin, antitoxin, and toxin-antitoxin complex have been more thoroughly investigated, little remains known about its transcription regulator PaaR2. Using a wide range of biophysical techniques (circular dichroism spectroscopy, size-exclusion chromatography-multiangle laser light scattering, dynamic light scattering, small-angle x-ray scattering, and native mass spectrometry), we demonstrate that PaaR2 mainly consists of α-helices and displays a concentration-dependent octameric build-up in solution and that this octamer contains a global shape that is significantly nonspherical. Thermal unfolding of PaaR2 is reversible and displays several transitions, suggesting a complex unfolding mechanism. The unfolding data obtained from spectroscopic and calorimetric methods were combined into a unifying thermodynamic model, which suggests a five-state unfolding trajectory. Furthermore, the model allows the calculation of a stability phase diagram, which shows that, under physiological conditions, PaaR2 mainly exists as a dimer that can swiftly oligomerize into an octamer depending on local protein concentrations. These findings, based on a thorough biophysical and thermodynamic analysis of PaaR2, may provide important insights into biological function such as DNA binding and transcriptional regulation.
|Alternate Journal||Biophys. J.|
|PubMed Central ID||PMC6486494|
Thermodynamic Stability of the Transcription Regulator PaaR2 from Escherichia coli O157:H7.