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- W2464662701 abstract "Flavohemoglobins (FlavoHbs) are a class of protein found in a large number of prokaryotic and eukaryotic microbes. These proteins are involved in NO signaling and detoxification through an NO dioxygenase mechanism. They are comprised of an N-terminal heme globin domain which allows for the binding of gaseous ligands and a C-terminal NADH/FADH binding domain which facilitates association of redox cofactors necessary for reduction of the bound ligand. This NO dioxygenase function is important in facilitating immune resistance by protecting the cell from nitrosative stress brought about by a host organism; as a result bacterial flavoHbs have recently been considered as targets for the development of new classes of antibiotics. In this study, photoacoustic calorimetry (PAC) and transient absorption spectroscopy (TA) have been used to characterize the energetics, structural dynamics, and kinetics of CO migration within flavoHbs from Ralstonia eutropha (FHP) and Staphylococcus aureus (HMPSa) with and without the addition of antibiotic azole compounds. We observe that CO escape from both proteins occurs within ∼50 ns of photodissociation. In Fhp, the ligand escape is associated with ΔH = 25.2±5.5 kcal mol-1 and ΔV = 20.6±1.2 mL mol-1 while in HMPSa ΔH = 73.0±5.9 kcal mol-1 and ΔV = 40.4±5.7 mL mol-1 was observed. These values indicated a distinct structural relaxation associated with ligand escape from both proteins. Multi-exponential kinetics observed for CO association to FHP and HMPSa suggest that multiple pathways for ligand entry to the heme binding pocket may exist in this protein. In addition, both proteins display somewhat temperature-dependent quantum yields for bimolecular rebinding of CO - approx. 0.65 for FHP and 0.32 for HMPSa at 20 °C." @default.
- W2464662701 created "2016-07-22" @default.
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- W2464662701 date "2016-02-01" @default.
- W2464662701 modified "2023-09-26" @default.
- W2464662701 title "Determination of Thermodynamic and Kinetic Parameters for Co Migration within Bacterial Flavohemoglobins of Ralstonia Eutropha and Staphylococcus Aureus" @default.
- W2464662701 doi "https://doi.org/10.1016/j.bpj.2015.11.1236" @default.
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