PELDOR/DEER: An Electron Paramagnetic Resonance Method to Study Membrane Proteins in Lipid Bilayers

Peter, Μ.; Bountra, Kiran; Beis, Konstantinos; Hagelueken, Gregor

doi:10.1007/978-1-0716-0724-4_15

Cited by 1 publication

(1 citation statement)

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“…EPR spectroscopy has emerged as an excellent tool for resolving such systems since it does not require crystallization and does not depend on protein size. EPR can be measured in buffer solution, and even a weak interaction between protein and DNA can be detected. − EPR’s strength lies in its sensitivity to both changes on an atomic level and nanoscale fluctuations. − In addition, EPR can measure distances between paramagnetic probes within the protein, and between proteins, up to 10.0 nm. − Sometimes the measurement of one distance (or a few) is sufficient to establish the plausibility of a mechanism or to validate a proposed structure. The most common means for obtaining nanoscale structural information is by pulsed electron double resonance (PELDOR), also commonly termed the double electron resonance experiment (DEER).…”

Section: Introductionmentioning

confidence: 99%

Revealing the DNA Binding Modes of CsoR by EPR Spectroscopy

Igbaria-Jaber,

Hofmann,

Gevorkyan-Airapetov

et al. 2023

ACS Omega

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In pathogens, a unique class of metalloregulator proteins, called gene regulatory proteins, sense specific metal ions that initiate gene transcription of proteins that export metal ions from the cell, thereby preventing toxicity and cell death. CsoR is a metalloregulator protein found in various bacterial systems that "sense" Cu(I) ions with high affinity. Upon copper binding, CsoR dissociates from the DNA promoter region, resulting in initiation of gene transcription. Crystal structures of CsoR in the presence and absence of Cu(I) from various bacterial systems have been reported, suggesting either a dimeric or tetrameric structure of these helical proteins. However, structural information about the CsoR-DNA complex is missing. Here, we applied electron paramagnetic resonance (EPR) spectroscopy to follow the conformational and dynamical changes that Mycobacterium tuberculosis CsoR undergoes upon DNA binding in solution. We showed that the quaternary structure is predominantly dimeric in solution, and only minor conformational and dynamical changes occur in the DNA bound state. Also, labeling of the unresolved C-terminus revealed no significant change in dynamics upon DNA binding. These observations are unique, since for other bacterial copper metalloregulators, such as the MerR and CopY families, major conformational changes were observed upon DNA binding, indicating a different mode of action for this protein family.

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