Role of Arginine 220 in the Oxygen Sensor FixL from Bradyrhizobium japonicum
| Autor: | Latifa Bouzhir-Sima, Tony A. Mattioli, Marten H. Vos, Laurent Kiger, Ursula Liebl, Véronique Balland, Michael C. Marden |
|---|---|
| Přispěvatelé: | CNRS, URA2096, Gif sur Yvette, F-91191, Centre National de la Recherche Scientifique (CNRS), Laboratoire d'optique et biosciences (LOB), École polytechnique (X)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Substitut du sang et pathologie moléculaire du globule rouge, Institut National de la Santé et de la Recherche Médicale (INSERM) |
| Rok vydání: | 2005 |
| Předmět: |
Hemeproteins
Models Molecular Time Factors Histidine Kinase Arginine Protein Conformation Ultraviolet Rays Electrons Heme Ligands Spectrum Analysis Raman 010402 general chemistry 01 natural sciences Biochemistry 03 medical and health sciences chemistry.chemical_compound Bacterial Proteins Spectroscopy Fourier Transform Infrared Bradyrhizobium Hypoxia Molecular Biology Histidine 030304 developmental biology Carbon Monoxide 0303 health sciences biology Histidine kinase Hydrogen Bonding DNA Cell Biology biology.organism_classification Protein Structure Tertiary 0104 chemical sciences Oxygen Kinetics Models Chemical chemistry Spectrophotometry Mutation Isoleucine Signal transduction Oxygen binding Signal Transduction Bradyrhizobium japonicum |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2005, 280, pp.15279-15288. ⟨10.1074/jbc.M413928200⟩ |
| ISSN: | 0021-9258 1083-351X |
| DOI: | 10.1074/jbc.m413928200 |
| Popis: | International audience; In the heme-based oxygen sensor protein FixL, conformational changes induced by oxygen binding to the heme sensor domain regulate the activity of a neighboring histidine kinase, eventually restricting expression of specific genes to hypoxic conditions. The conserved arginine 220 residue is suggested to play a key role in the signal transduction mechanism. To obtain detailed insights into the role of this residue, we replaced Arg220 by histidine (R220H), glutamine (R220Q), glutamate (R220E), and isoleucine (R220I) in the heme domain FixLH from Bradyrhizobium japonicum. These mutations resulted in dramatic changes in the O2 affinity with Kd values in the order R220I < R220Q < wild type < R220H. For the R220H and R220Q mutants, residue 220 interacts with the bound O2 or CO ligands, as seen by resonance Raman spectroscopy. For the oxy-adducts, this H-bond modifies the π acidity of the O2 ligand, and its strength is correlated with the back-bonding-sensitive ν4 frequency, the koff value for O2 dissociation, and heme core-size conformational changes. This effect is especially strong for the wild-type protein where Arg220 is, in addition, positively charged. These observations strongly suggest that neither strong ligand fixation nor the displacement of residue 220 into the heme distal pocket are solely responsible for the reported heme conformational changes associated with kinase activity regulation, but that a significant decrease of the heme π* electron density because of strong back-bonding toward the oxygen ligand also plays a key role. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|