Dynamics of the Heme-binding Bacterial Gas-sensing Dissimilative Nitrate Respiration Regulator (DNR) and Activation Barriers for Ligand Binding and Escape.

Laura Lobato 1 Latifa Bouzhir-Sima 1 Taku Yamashita Michael T Wilson 2 Marten H Vos 1 Ursula Liebl 1
1 LOB - Laboratoire d'optique et biosciences
2 Department of Sports Medicine
Abstract : DNR (dissimilative nitrate respiration regulator) is a heme-binding transcription factor that is involved in the regulation of denitrification in Pseudomonas aeruginosa. In the ferrous deoxy state, the heme is 6-coordinate; external NO and CO can replace an internal ligand. Using fluorescence anisotropy, we show that high-affinity sequence-specific DNA binding occurs only when the heme is nitrosylated, consistent with the proposed function of DNR as NO sensor and transcriptional activator. This role is moreover supported by the NO "trapping" properties revealed by ultrafast spectroscopy that are similar to those of other heme-based NO sensor proteins. Dissociated CO-heme pairs rebind in an essentially barrierless way. This process competes with migration out of the heme pocket. The latter process is thermally activated (Ea ∼7 kJ/mol). This result is compared with other heme proteins, including the homologous CO sensor/transcription factor CooA, variants of the 5-coordinate mycobacterial sensor DosT and the electron transfer protein cytochrome c. This comparison indicates that thermal activation of ligand escape from the heme pocket is specific for systems where an external ligand replaces an internal one. The origin of this finding and possible implications are discussed.
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Article dans une revue
Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2014, 289 (38), pp.26514-24. 〈10.1074/jbc.M114.571398 〉
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Contributeur : Denis Roura <>
Soumis le : jeudi 30 octobre 2014 - 20:30:12
Dernière modification le : jeudi 10 mai 2018 - 02:08:18

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Laura Lobato, Latifa Bouzhir-Sima, Taku Yamashita, Michael T Wilson, Marten H Vos, et al.. Dynamics of the Heme-binding Bacterial Gas-sensing Dissimilative Nitrate Respiration Regulator (DNR) and Activation Barriers for Ligand Binding and Escape.. Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2014, 289 (38), pp.26514-24. 〈10.1074/jbc.M114.571398 〉. 〈hal-01079002〉

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