Ferric Uptake Regulator (Fur) Reversibly Binds a [2Fe-2S] Cluster to Sense Intracellular Iron Homeostasis in Escherichia Coli

The ferric uptake regulator (Fur) is a global transcription factor that regulates intracellular iron homeostasis in bacteria. The current hypothesis states that when the intracellular “free” iron concentration is elevated, Fur binds ferrous iron, and the iron-bound Fur represses the genes encoding for iron uptake systems and stimulates the genes encoding for iron storage proteins. However, the “iron-bound” Fur has never been isolated from any bacteria. Here we report that the Escherichia coli Fur has a bright red color when expressed in E. coli mutant cells containing an elevated intracellular free iron content because of deletion of the iron–sulfur cluster assembly proteins IscA and SufA. The acid-labile iron and sulfide content analyses in conjunction with the EPR and Mössbauer spectroscopy measurements and the site-directed mutagenesis studies show that the red Fur protein binds a [2Fe-2S] cluster via conserved cysteine residues. The occupancy of the [2Fe-2S] cluster in Fur protein is ∼31% in the E. coli iscA/sufA mutant cells and is decreased to ∼4% in WT E. coli cells. Depletion of the intracellular free iron content using the membrane-permeable iron chelator 2,2´-dipyridyl effectively removes the [2Fe-2S] cluster from Fur in E. coli cells, suggesting that Fur senses the intracellular free iron content via reversible binding of a [2Fe-2S] cluster. The binding of the [2Fe-2S] cluster in Fur appears to be highly conserved, because the Fur homolog from Hemophilus influenzae expressed in E. coli cells also reversibly binds a [2Fe-2S] cluster to sense intracellular iron homeostasis

Ferric uptake regulator (Fur) reversibly binds a [2Fe-2S] cluster to sense intracellular iron homeostasis in escherichia coli

© 2020 Fontenot et al. The ferric uptake regulator (Fur) is a global transcription factor that regulates intracellular iron homeostasis in bacteria. The current hypothesis states that when the intracellular “free” iron concentration is elevated, Fur binds ferrous iron, and the iron-bound Fur represses the genes encoding for iron uptake systems and stimulates the genes encoding for iron storage proteins. However, the “iron-bound” Fur has never been isolated from any bacteria. Here we report that the Escherichia coli Fur has a bright red color when expressed in E. coli mutant cells containing an elevated intracellular free iron content because of deletion of the iron–sulfur cluster assembly proteins IscA and SufA. The acid-labile iron and sulfide content analyses in conjunction with the EPR and Mössbauer spectroscopy measurements and the site-directed mutagenesis studies show that the red Fur protein binds a [2Fe-2S] cluster via conserved cysteine residues. The occupancy of the [2Fe-2S] cluster in Fur protein is ~31% in the E. coli iscA/sufA mutant cells and is decreased to ~4% in WT E. coli cells. Depletion of the intracellular free iron content using the membrane-permeable iron chelator 2,2´-dipyridyl effectively removes the [2Fe-2S] cluster from Fur in E. coli cells, suggesting that Fur senses the intracellular free iron content via reversible binding of a [2Fe-2S] cluster. The binding of the [2Fe-2S] cluster in Fur appears to be highly conserved, because the Fur homolog from Hemophilus influenzae expressed in E. coli cells also reversibly binds a [2Fe-2S] cluster to sense intracellular iron homeostasis

Molecular and Electronic Structures and Single-Molecule Magnet Behavior of Tris(thioether)-Iron Complexes Containing Redox-Active α-Diimine Ligands

Incorporating radical ligands into metal complexes is one of the emerging trends in the design of single-molecule magnets (SMMs). While significant effort has been expended to generate multinuclear transition metal-based SMMs with bridging radical ligands, less attention has been paid to mononuclear transition metal–radical SMMs. Herein, we describe the first α-diiminato radical-containing mononuclear transition metal SMM, namely, [κ2-PhTt tBu]­Fe­(AdNCHCHNAd) (1), and its analogue [κ2-PhTt tBu]­Fe­(CyNCHCHNCy) (2) (PhTt tBu = phenyltris­(tert-butylthiomethyl)­borate, Ad = adamantyl, and Cy = cyclohexyl). 1 and 2 feature nearly identical geometric and electronic structures, as shown by X-ray crystallography and electronic absorption spectroscopy. A more detailed description of the electronic structure of 1 was obtained through EPR and Mössbauer spectroscopies, SQUID magnetometry, and DFT, TD-DFT, and CAS calculations. 1 and 2 are best described as high-spin iron­(II) complexes with antiferromagnetically coupled α-diiminato radical ligands. A strong magnetic exchange coupling between the iron­(II) ion and the ligand radical was confirmed in 1, with an estimated coupling constant J \u3c −250 cm–1 (J = −657 cm–1, DFT). Calibrated CAS calculations revealed that the ground-state Fe­(II)−α-diiminato radical configuration has significant ionic contributions, which are weighted specifically toward the Fe­(I)-neutral α-diimine species. Experimental data and theoretical calculations also suggest that 1 possesses an easy-axis anisotropy, with an axial zero-field splitting parameter D in the range from −4 to–1 cm–1. Finally, dynamic magnetic studies show that 1 exhibits slow magnetic relaxation behavior with an energy barrier close to the theoretical maximum, 2|D|. These results demonstrate that incorporating strongly coupled α-diiminato radicals into mononuclear transition metal complexes can be an effective strategy to prepare SMMs

Redox active iron nitrosyl units in proton reduction electrocatalysis

Base metal, molecular catalysts for the fundamental process of conversion of protons and electrons to dihydrogen, remain a substantial synthetic goal related to a sustainable energy future. Here we report a diiron complex with bridging thiolates in the butterfly shape of the 2Fe2S core of the [FeFe]-hydrogenase active site but with nitrosyl rather than carbonyl or cyanide ligands. This binuclear [(NO)Fe(N_2S_2)Fe(NO)_2]+ complex maintains structural integrity in two redox levels; it consists of a (N_2S_2)Fe(NO) complex (N_2S_2=N,N′-bis(2-mercaptoethyl)-1,4-diazacycloheptane) that serves as redox active metallodithiolato bidentate ligand to a redox active dinitrosyl iron unit, Fe(NO)_2. Experimental and theoretical methods demonstrate the accommodation of redox levels in both components of the complex, each involving electronically versatile nitrosyl ligands. An interplay of orbital mixing between the Fe(NO) and Fe(NO)_2 sites and within the iron nitrosyl bonds in each moiety is revealed, accounting for the interactions that facilitate electron uptake, storage and proton reduction

2015/16 seasonal vaccine effectiveness against hospitalisation with influenza a(H1N1)pdm09 and B among elderly people in Europe: Results from the I-MOVE+ project

We conducted a multicentre test-negative caseâ\u80\u93control study in 27 hospitals of 11 European countries to measure 2015/16 influenza vaccine effectiveness (IVE) against hospitalised influenza A(H1N1)pdm09 and B among people aged â\u89¥ 65 years. Patients swabbed within 7 days after onset of symptoms compatible with severe acute respiratory infection were included. Information on demographics, vaccination and underlying conditions was collected. Using logistic regression, we measured IVE adjusted for potential confounders. We included 355 influenza A(H1N1)pdm09 cases, 110 influenza B cases, and 1,274 controls. Adjusted IVE against influenza A(H1N1)pdm09 was 42% (95% confidence interval (CI): 22 to 57). It was 59% (95% CI: 23 to 78), 48% (95% CI: 5 to 71), 43% (95% CI: 8 to 65) and 39% (95% CI: 7 to 60) in patients with diabetes mellitus, cancer, lung and heart disease, respectively. Adjusted IVE against influenza B was 52% (95% CI: 24 to 70). It was 62% (95% CI: 5 to 85), 60% (95% CI: 18 to 80) and 36% (95% CI: -23 to 67) in patients with diabetes mellitus, lung and heart disease, respectively. 2015/16 IVE estimates against hospitalised influenza in elderly people was moderate against influenza A(H1N1)pdm09 and B, including among those with diabetes mellitus, cancer, lung or heart diseases

Moessbauer studies of model complexes for the H-cluster in [Fe-Fe]-hydrogenases: Electronic structure of mixed-valence complexes containing low-spin Fe(I)

[[abstract]]Hydrogenases are enzymes that catalyze the reversible conversion of protons to mol. hydrogen. The active site of the [Fe-Fe]-hydrogenases (the H-cluster) can provide the required potential for this reaction using iron in low oxidn. states. Since it has been recognized that the H-cluster may contain low-spin Fe(I), monovalent iron has become a focus of bio-organometallic synthetic chem. In spite of its intriguing implication in hydrogenogenesis, the electronic structure of low-spin Fe(I) is unknown. The high asymmetry of the dinuclear site in the H-cluster stabilizes the elusive Fe(I)Fe(II...[[conferencetype]]國際[[conferencedate]]20120819~20120823[[booktype]]紙本[[iscallforpapers]]Y[[conferencelocation]]Philadelphia, PA, United State

Moessbauer studies of mixed-valence Fe(II)Fe(I) and Fe(I)Fe(I) model complexes illustrating states of the H-cluster in [Fe-Fe]-hydrogenases

[[abstract]]Hydrogenases are iron enzymes that catalyze the reversible conversion of protons to mol. hydrogen. [Fe-Fe] hydrogenases are known for efficient prodn. of H2, at their active site (the H-cluster) thus their structure and mechanism constitute both inspiration and a target for catalyst design. Since it has been recognized that the H-cluster contains low-spin Fe(I), monovalent iron has become a focus of bio-organometallic synthetic chem. The high asymmetry of the dinuclear site stabilizes the elusive Fe(I)Fe(II) oxidized state, Hox. Oxidn. of asym. complexes [PMe3(CO)2FeI(μ-pdt)FeI(CO)2IMes] ...[[conferencetype]]國際[[conferencedate]]20120819~20120823[[booktype]]紙本[[iscallforpapers]]Y[[conferencelocation]]Philadelphia, PA, United State