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Thermodynamic stability of the transcription regulator PaaR2 from Escherichia coli O157:H7. / De Bruyn, Pieter; Hadzi, San; Vandervelde, Alexandra; Konijnenberg, Albert; Prolič Kalinšek, Maruša; Sterckx, Yann; Sobott, Frank; Lah, Jurij; Van Melderen, Laurence; Loris, Remy.

In: Biophysical Journal, Vol. 116, No. 8, 23.04.2019, p. 1420-1431.

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De Bruyn, Pieter ; Hadzi, San ; Vandervelde, Alexandra ; Konijnenberg, Albert ; Prolič Kalinšek, Maruša ; Sterckx, Yann ; Sobott, Frank ; Lah, Jurij ; Van Melderen, Laurence ; Loris, Remy. / Thermodynamic stability of the transcription regulator PaaR2 from Escherichia coli O157:H7. In: Biophysical Journal. 2019 ; Vol. 116, No. 8. pp. 1420-1431.

BibTeX

@article{8aa49bee89a94e2eb3068336c7bf1f0b,
title = "Thermodynamic stability of the transcription regulator PaaR2 from Escherichia coli O157:H7",
abstract = "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.",
keywords = "Toxin-antitoxin module, Transcription factors, Molecular biophysics, Protein chemistry",
author = "{De Bruyn}, Pieter and San Hadzi and Alexandra Vandervelde and Albert Konijnenberg and {Prolič Kalinšek}, Maruša and Yann Sterckx and Frank Sobott and Jurij Lah and {Van Melderen}, Laurence and Remy Loris",
note = "Copyright {\circledC} 2019 Biophysical Society. Published by Elsevier Inc. All rights reserved.",
year = "2019",
month = "4",
day = "23",
doi = "https://doi.org/10.1016/j.bpj.2019.03.015",
language = "English",
volume = "116",
pages = "1420--1431",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Biophysical Society",
number = "8",

}

RIS

TY - JOUR

T1 - Thermodynamic stability of the transcription regulator PaaR2 from Escherichia coli O157:H7

AU - De Bruyn, Pieter

AU - Hadzi, San

AU - Vandervelde, Alexandra

AU - Konijnenberg, Albert

AU - Prolič Kalinšek, Maruša

AU - Sterckx, Yann

AU - Sobott, Frank

AU - Lah, Jurij

AU - Van Melderen, Laurence

AU - Loris, Remy

N1 - Copyright © 2019 Biophysical Society. Published by Elsevier Inc. All rights reserved.

PY - 2019/4/23

Y1 - 2019/4/23

N2 - 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.

AB - 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.

KW - Toxin-antitoxin module

KW - Transcription factors

KW - Molecular biophysics

KW - Protein chemistry

UR - http://www.scopus.com/inward/record.url?scp=85063982820&partnerID=8YFLogxK

U2 - https://doi.org/10.1016/j.bpj.2019.03.015

DO - https://doi.org/10.1016/j.bpj.2019.03.015

M3 - Article

VL - 116

SP - 1420

EP - 1431

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 8

ER -

ID: 45236901