In vitro coordination of Fe-protoheme with amyloid β is non-specific and exhibits multiple equilibria.

In addition to copper and zinc, heme is thought to play a role in Alzheimer's disease and its metabolism is strongly affected during the course of this disease. Amyloid β, the peptide associated with Alzheimer's disease, was shown to bind heme in vitro with potential catalytic activity linked to oxidative stress. To date, there is no direct determination of the structure of this complex.

Conformational H-bonding modulation of the iron active site cysteine ligand of superoxide reductase: absorption and resonance Raman studies.

Superoxide reductases (SORs) are mononuclear non-heme iron enzymes involved in superoxide radical detoxification in some microorganisms. Their atypical active site is made of an iron atom pentacoordinated by four equatorial nitrogen atoms from histidine residues and one axial sulfur atom from a cysteinate residue, which plays a central role in catalysis. In most SORs, the residue immediately following the cysteinate ligand is an asparagine, which belongs to the second coordination sphere and is expected to have a critical influence on the properties of the active site.

Modulation of the flavin-protein interactions in NADH peroxidase and mercuric ion reductase: a resonance Raman study.

NADH peroxidase (Npx) and mercuric ion reductase (MerA) are flavoproteins belonging to the pyridine nucleotide:disulfide oxidoreductases (PNDO) and catalyzing the reduction of toxic substrates, i.e., hydrogen peroxide and mercuric ion, respectively.

Formation of high-valent iron-oxo species in superoxide reductase: characterization by resonance Raman spectroscopy.

Superoxide reductase (SOR), a non-heme mononuclear iron protein that is involved in superoxide detoxification in microorganisms, can be used as an unprecedented model to study the mechanisms of O2 activation and of the formation of high-valent iron-oxo species in metalloenzymes. By using resonance Raman spectroscopy, it was shown that the mutation of two residues in the second coordination sphere of the SOR iron active site, K48 and I118, led to the formation of a high-valent iron-oxo species when the mutant proteins were reacted with H2O2. These data demonstrate that these residues in the second coordination sphere tightly control the evolution and the cleavage of the O-O bond of the ferric iron hydroperoxide intermediate that is formed in the SOR active site.

Hydrogen bonding to the cysteine ligand of superoxide reductase: acid-base control of the reaction intermediates.

Superoxide reductase (SOR) is a non-heme iron metalloenzyme that detoxifies superoxide radical in microorganisms. Its active site consists of an unusual non-heme Fe(2+) center in a [His4Cys1] square pyramidal pentacoordination, with the axial cysteine ligand proposed to be an essential feature in catalysis. Two NH peptide groups from isoleucine 118 and histidine 119 establish hydrogen bonds involving the sulfur ligand (Desulfoarculus baarsii SOR numbering).

Leghemoglobin green derivatives with nitrated hemes evidence production of highly reactive nitrogen species during aging of legume nodules.

Globins constitute a superfamily of proteins widespread in all kingdoms of life, where they fulfill multiple functions, such as efficient O(2) transport and modulation of nitric oxide bioactivity. In plants, the most abundant Hbs are the symbiotic leghemoglobins (Lbs) that scavenge O(2) and facilitate its diffusion to the N(2)-fixing bacteroids in nodules. The biosynthesis of Lbs during nodule formation has been studied in detail, whereas little is known about the green derivatives of Lbs generated during nodule senescence.

Redox effects on the coordination geometry and heme conformation of bis(N-methylimidazole) complexes of superstructured Fe-porphyrins. A spectroscopic study.

Electronic absorption, electron paramagnetic resonance (EPR), and Soret-excited resonance Raman (RR) spectra are reported for bis(N-alkylimidazole) complexes of various iron(III)-"basket-handle" (Fe(III)BHP(+)) and "picket-fence" (Fe(III)PFP(+)) porphyrins in methylene chloride. The Fe(III)BHP(+) derivatives consist of four cross-trans (CT) and two adjacent-cis (AC) -linked in which the composition and the length of the handles are variable (CT Fe(III)[(C(11)Im)(2)(+)], CT and AC Fe(III)[((C(4))(2)phi)(2)](+), CT Fe(III)[((C(3))(2)phi)(C(12))](+), CT and AC Fe(III)[((C(3))(2)phi)(2)](+)). The meso-alphaalpha betabeta and meso-alphabeta alphabeta atropisomers of Fe(III)-tetrakis(o-pivalamidophenyl)-porphyrins represents the Fe(III)PFP(+) derivatives (Fe(III)alphaalpha betabeta-T(piv)PP(+) and Fe(III)alphabeta alphabeta-T(piv)PP(+), respectively).

Low-frequency heme, iron-ligand, and ligand modes of imidazole and imidazolate complexes of iron protoporphyrin and microperoxidase in aqueous solution. An analysis by far-infrared difference spectroscopy.

FTIR difference spectroscopy, notably in the far-IR domain, is appealing for the analysis of hemoproteins, since it permits us to directly probe the properties of the heme and its ligands but also those of aminoacids remote from the heme. We recently set a thin path-length electrochemical cell with diamond windows allowing the far-IR analysis of proteins in aqueous solutions using FTIR difference spectroscopy (Berthomieu, C,; Marboutin, L.; Dupeyrat, F.

Ultrafast heme-residue bond formation in six-coordinate heme proteins: implications for functional ligand exchange.

A survey is presented of picosecond kinetics of heme-residue bond formation after photolysis of histidine, methionine, or cysteine, in a broad range of ferrous six-coordinate heme proteins. These include human neuroglobin, a bacterial heme-binding superoxide dismutase (SOD), plant cytochrome b 559, the insect nuclear receptor E75, horse heart cytochrome c and the heme domain of the bacterial sensor protein Dos. We demonstrate that the fastest and dominant phase of binding of amino acid residues to domed heme invariably takes place with a time constant in the narrow range of 5-7 ps.

Interaction of glutathione reductase with heavy metal: the binding of Hg(II) or Cd(II) to the reduced enzyme affects both the redox dithiol pair and the flavin.

To determine the inhibition mechanism of yeast glutathione reductase (GR) by heavy metal, we have compared the electronic absorption and resonance Raman (RR) spectra of the enzyme in its oxidized (Eox) and two-electron reduced (EH2) forms, in the absence and the presence of Hg(II) or Cd(II). The spectral data clearly show a redox dependence of the metal binding. The metal ions do not affect the absorption and RR spectra of Eox.

Spectroscopic characterization of hydroxide and aqua complexes of Fe(II)-protoheme, structural models for the axial coordination of the atypical heme of membrane cytochrome b6f complexes.

Electronic absorption and resonance Raman (RR) spectra are reported for hydroxide and aqua complexes of iron(II)-protoporphyrin IX (Fe(II)PP) respectively formed in alkaline and neutral aqueous solutions. These compounds with weak axial ligand(s) represent a biomimetic approach of the unusual coordination of the atypical heme c(i) of membrane cytochrome b6f complexes. Absorption spectra and spectrophotometric titrations show that Fe(II)PP in alkaline aqueous cetyltrimethylammonium bromide (CTABr) binds one hydroxide ion, forming a five-coordinated high-spin (HS) complex.

Biochemical and spectroscopic characterization of the covalent binding of heme to cytochrome b6.

The three-dimensional structure of the cytochrome b(6)f complex disclosed the unexpected presence of a new heme c(i) [Stroebel, D., Choquet, Y., Popot, J.

Nonplanar distortions of bis-base low-spin iron(II)-porphyrinates: absorption and resonance Raman investigations of cross-trans-linked iron(II)-basket-handle porphyrin complexes.

Electronic absorption and Soret-excited resonance Raman (RR) spectra are reported for bis-N-alkylimidazole and bis-pyridine complexes of various cross-trans-linked iron(II)-"basket-handle" porphyrins (Fe(II)-BHP) in methylene chloride. These compounds enable us to characterize the spectroscopic properties of ruffled six-coordinated low-spin Fe(II)-porphyrin complexes. The visible absorption spectra show that the Q and B bands are progressively red-shifted when the handles are shortened and/or when the steric hindrance of the axial ligands is increased.

Absorption and resonance Raman investigations of ligand rotation and nonplanar heme distortion in bis-base low-spin iron(II)-tetrakis(o-pivalamidophenyl)porphyrin complexes.

The absorption and resonance Raman (RR) spectra of the bis-N-methylimidazole, bis-1,5-dicyclohexylimidazole, and bis-pyridine complexes of the meso-alphaalphabetabeta and meso-alphabetaalphabeta atropisomers of Fe(II)-tetrakis(o-pivalamidophenyl)porphyrins (Fe(II)TpivPP) were obtained in methylene chloride. The different spatial arrangements of the o-pivalamide pickets in these two Fe(II)TpivPP compounds are expected to control the absolute and relative positions of the axial ligand rings with respect to the Fe-N(pyrrole) bonds. In particular, the spectroscopic data obtained for the bis-N-methylimidazole and bis-dicyclohexylimidazole complexes of the Fe(II)[alphabetaalphabeta-TpivPP] derivative showed the most important differences.

Structural and EPR characterization of the soluble form of cytochrome c-550 and of the psbV2 gene product from the cyanobacterium Thermosynechococcus elongatus.

First, the crystal structure of cytochrome c-550 (the psbV1 gene product) from the thermophilic cyanobacterium Thermosynechococcus elongatus has been determined to a resolution of 1.8 A. A comparison of the T.

Two modes of binding of N-hydroxyguanidines to NO synthases: first evidence for the formation of iron-N-hydroxyguanidine complexes and key role of tetrahydrobiopterin in determining the binding mode.

The interaction of various N-alkyl- and N-aryl-N'-hydroxyguanidines with recombinant NOS containing or not containing tetrahydrobiopterin (BH(4)) was studied by visible, electronic paramagnetic resonance (EPR), and resonance Raman (RR) spectroscopy. N-Hydroxyguanidines interact with the oxygenase domain of BH(4)-free inducible NOS (BH(4)-free iNOS(oxy)), depending on the nature of their substituent, with formation of two types of complexes that are characterized by peaks around 395 (type I) and 438 nm (type II') during difference visible spectroscopy. The complex formed between BH(4)-free iNOS(oxy) and N-benzyl-N'-hydroxyguanidine 1 (type II') exhibited a Soret peak at 430 nm, EPR signals at g = 1.

Electrostatic control of the isoalloxazine environment in the two-electron reduced states of yeast glutathione reductase.

The resonance Raman spectra of the oxidized and two-electron reduced forms of yeast glutathione reductase are reported. The spectra of the oxidized enzyme indicate a low electron density for the isoalloxazine ring. As far as the two-electron reduced species are concerned, the spectral comparison of the NADPH-reduced enzyme with the glutathione- or dithiothreitol-reduced enzyme shows significant frequency differences for the flavin bands II, III, and VII.