Light-induced EPR spectra of reaction centers fromRhodobacter sphaeroides at 80 K: Evidence for reduction of QB by B branch electron transfer in native reaction centers

Photosynthetic reaction centers (RCs) fromRhodobacter sphaeroides capture solar energy by electron transfer from primary donor D to quinone acceptor Q_B through the active A branch of electron acceptors. The light-induced electron paramagnetic resonance (EPR) spectrum from native RCs that had Fe²⁺ replaced by Zn²⁺ was investigated at cryogenic temperature (80 K, 35 GHz). In addition to the light-induced signal due to the formation of D^+.Q _A ^?. observed previously, a small fraction (ca. 5%) of the signal displayed very different characteristics. The signal was absent in RCs in which the Q_B was displaced by the inhibitor stigmatellin. Its decay time (τ=6 s) was the same as observed for D^+.Q _B ^?. in mutant RCs lacking Q_A, which is significantly slower than for D^+.Q _A ^?. (τ=30 ms). Its EPR spectrum was identical to that of D^+.Q _B ^?. . The quantum efficiency for forming the major component of the signal was the same as that found for mutant RCs lacking Q_A (?=0.2%) and was temperature independent. These results are explained by direct photochemical reduction of Q_B via B branch electron transfer in a small fraction of native RCs.     

Influence of heme periphery upon electronic structúre and Mössbauer spectra parameters of hemoglobin

Quantum-chemical analysis of electronic structure peculiarities and mossbauer spectra parameters was performed for penta-coordinated complex of ferro-protoporphyrin with imidazole (Fe(+2)PPIm). Peripheral substitutes (−CH₃, −C₂H₃, −C₂H₄COOH) introduced in the porphyrin macrocycle simulated real chemical structure of protoporphyrin (PP) in heme group of Hb. Calculations displayed that doubly occupied molecular orbitals (MO) of the peripheral substitutes (−CH=CH₂ and −CH₂−CH₂−COOH) always appeared near the occupat ion border. The orientation of vinyl fragment have the essential influence upon Fe₅ dorbital populations and quadrupole splitting_AE_Q for⁵B₁ and⁵B₂ terms. The values of isomer shift were insensitive to that modification of fragment orientation.     

The transient EPR spectra and spin dynamics of coupled three-spin systems in photosynthetic reaction centres

The concept of introducing an additional, stable paramagnetic species into photosynthetic reaction centres to increase the information content of their spin polarized transient EPR spectra is investigated theoretically. The light-induced electron transfer in such systems generates a series of coupled three-spin states consisting of sequential photoinduced radical pairs coupled to the stable spin which acts as an “observer”. The spin polarized transient EPR spectra are investigated using the coupled three-spin system P⁺I⁻Q⁻ _A in pre-reduced bacterial reaction centres as a specific example which has been studied experimentally. The evolution of the spin system and the spin polarized EPR spectra of P⁺I⁻Q⁻ _A and Q⁻ _A following recombination of the radical pair (P = primary donor, I = primary acceptor, Q_A = quinone acceptor) are calculated numerically by solving the equations of motion for the density matrix. The net polarization of the observer spin is also calculated analytically by perturbation theory for the case of a single, short-lived, charge-separated state. The result bears a close resemblance to the chemically induced nuclear polarization (CIDNP) generated in photolysis reactions in which a nuclear spin plays the role of the observer interacting with the radical pair intermediates. However, because the Zeeman frequencies of the three electron spins involved are usually quite similar, the polarization of the electron observer spin in strong magnetic fields can reflect features of the CIDNP effect in both, high and low magnetic fields. The dependence of the quinone spin polarization on the exchange couplings in the three-spin system is investigated by numerical simulations, and it is shown that the observed emissive polarization pattern is compatible with either sign, positive or negative, for a range of exchange couplings, J_PI, in the primary pair. The microwave frequency and orientation dependence of the spectra are discussed as two of several possible criteria for determining the sign of J_PI.     

Flexible-to-semiflexible chain crossover on the pressure-area isotherm of a lipid bilayer

We find theoretically that competition between ∼K _f q ⁴ and ∼Qq ² terms in the Fourier-transformed conformational energy of a single-lipid chain, in combination with interchain entropic repulsion in the hydrophobic part of the lipid (bi)layer, may cause a crossover on the bilayer pressure-area isotherm P(A)∼(A−A ₀)^−α. The crossover manifests itself in the transition from α = 5/3 to α = 3. Our microscopic model represents a single-lipid molecule as a worm-like chain with a finite irreducible cross-section area A ₀, a flexural rigidity K _f , and a stretching modulus Q in a parabolic potential with the self-consistent curvature B(A) formed by entropic interactions between hydrocarbon chains in the lipid layer. The crossover area A* obeys the relation Q/√K _f B(A*) ≈ 2. We predict a peculiar possibility of deducing the effective elastic moduli K _f and Q of an individual hydrocarbon chain from the analysis of the isotherm with such a crossover. Also calculated is the crossover-related behavior of the area compressibility modulus K _A , the equilibrium area per lipid A _t , and the chain order parameter S(θ). The text was submitted by the authors in English.     

Mössbauer study of ultrafine amorphous Fe−B alloys

The chemical composition of ultrafine amorphous Fe−B powders prepared by a chemical reduction depends on the mixed molar ratio of KBH₄ to Fe ions. We propose the following reaction processes for the formation of ultrafine Fe−B powders: (1) 4Fe²⁺+2BH₄₋+6OH⁻→2Fe₂B+6H₂O+H₂ and (2) 4Fe²⁺+2BH₄₋+7OH⁻→2Fe₃B+Fe+BO₂+5H₂O+5/2H₂.     

The primary and secondary acceptors in bacterial photosynthesis III. Characterization of the quinone radicals Q A − ⋅ and Q B − ⋅ by EPR and ENDOR

Electron paramagnetic resonance (EPR) and electron-nuclear double resonance (ENDOR) techniques were used to investigate the electronic structure of the primary (Q _A ^?? ) and secondary (Q _B ^?? ) ubiquinone electron acceptors in reaction centers (RCs) of the photosynthetic bacteriumRhodobacter sphaeroides. To reduce the EPR linewidth, the high-spin Fe²⁺ present in native RCs was replaced by diamagnetic Zn²⁺. Experiments were performed both on frozen solutions and single crystals at microwave frequencies of 9, 35 and 94 GHz. Differences in the EPR/ENDOR spectra were observed for Q _A ^?? and Q _B ^?? , which are attributed to different environments of the quinones in the RC. The differences exhibited themselves in: (i) the g-tensors, (ii) the¹⁷O and¹³C hyperfine coupling (hfc) constants of the quinones labeled at the carbonyl group, (iii) the¹H-hfcs of the quinone ring and (iv) the exchangeable protons hydrogen bonded to the carbonyl oxygens. From these results and from H/D exchange experiments, the following conclusions were drawn: both Q _A ^?? and Q _B ^?? have at least two hydrogen bonds of different strengths to the carbonyl oxygens. The hydrogen bonds for Q _A ^?? are stronger and more asymmetric than for Q _B ^?? . For Q _A ^?? the stronger bond (to O₄) was assigned to His(M219) and the weaker (to O₁) to Ala(M260). For Q _B ^?? the stronger bond (to O₄) was assigned to His(L190), with several weaker bonds (to O₁) to Ser(L223), Ile(224) and Gly(L225). From the temperature dependence of the hfcs of the exchangeable protons some dynamic properties of the RC were deduced. Hfcs with more distant nitrogens were observed by electron spin echo envelope modulation (ESEEM). For Q _A ^?? they were assigned to N_δ of His(M219) and to the peptide backbone nitrogen of Ala(M260) and for Q _B ^?? to N_δ of His(L190). These interactions indicate the extent of the electron wave function, which is important for the understanding of the electron transfer mechanism. Based on the magnetic resonance results, the function of the quinone acceptors in the reaction center is discussed.     

Model for primary electron transfer and coupling of electronic states at reaction centers of purple bacteria

A detailed derivation is presented for relations making it possible to describe the effect of temperature on the halfwidth of the P960 and P870 absorption bands and also on the electron transfer (ET) rate at reaction centers (RCs) of the purple bacteria Rps. viridis and Rb. sphaeroides. Primary electron transfer is considered as a resonant nonradiative transition between P* and P⁺B _L ⁻ states (where P is a special pair, B_L is an additional bacteriochlorophyll in the L branch of the reaction center). It has been shown that the vibrational h_α mode with frequency 130–150 cm⁻¹ controls primary electron transfer. It has been found that the matrix element of the electronic transition between the states P* and P⁺B _L ⁻ is equal to 12.7 ± 0.9 and 12.0 ± 1.2 cm⁻¹ for Rps. viridis and Rb. sphaeroides respectively. The mechanism is discussed for electron transport from P* and B_L and then to bacteriopheophytin H_L. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 73, No. 3, pp. 294–303, May–June, 2006.     

EPR of trivalent iron ions in a LiCaAlF6 crystal

The orientational dependences of the EPR spectra of Fe³⁺-doped LiCaAlF₆ single crystals (space group P31c, Z=2), grown at the Laboratory of Magnetic Radio Spectroscopy at Kazan’ State University, have been investigated in detail. The spectrum is described by a trigonal spin Hamiltonian with the following parameters: B ₂₀=40.072×10⁻⁴ cm⁻¹, B ₄₀=−5.799×10⁻⁴ cm⁻¹, B ₄₃=−4.281×10⁻⁴ cm⁻¹, A _s=24.33±1, A _p=6.13±1, g _∥=g _⊥=2.00217±0.0003. A theoretical calculation of the hyperfine structure parameters shows that they are described quite well when allowance is made for the overlapping of the wave functions of the paramagnetic center and the ligands (F⁻). Fiz. Tverd. Tela (St. Petersburg) 39, 488–490 (March 1997)     

13C−1H heteronuclear dipolar correlation studies of the hydrogen bonding of the quinones inRhodobacter sphaeroides R26 reaction centers

The photosynthetic reaction center (RC) of the photosynthetic bacteriumRhodobacter sphaeroides R26 contains two quinones, Q_A and Q_B. Solid-state heteronuclear (¹H?¹³C) dipolar correlation spectroscopy has been used to study the binding of the quinones in the ground state for RCs reconstituted with l-¹³C ubiquinone-10. Lee-Goldburg cross-polarization buildup curves are recorded to determine distancesr _CH between the l-¹³C carbon labels and the protons involved in the polarization transfer. The l-¹³C of both Q_A and Q_B have intermolecular correlations with protons that resonate downfield, in the region of hydrogen-bonding protons. The distances between the carbon labels and the correlated protons are short, 0.21±0.01 nm. Hence the nuclear magnetic resonance provides evidence for strong hydrogen-bonding interactions at the l-C=O of both Q_A and Q_B for RCs in the ground state. The environment of the l-¹³C of the Q_B is structurally heterogeneous compared to that of the Q_A. The data can be reconciled with a strong H-bonding interaction of the l-C=O of Q_A with Ala M260 NH, and with complex hydrogen bonding involving NH of Ile-L224 and of Gly-L225, and possibly the Ser-L223 hydroxyl group of the l-C=O of the Q_B, in the proximal site.     

The primary and secondary acceptors in bacterial photosynthesis: II. The structure of the Fe2+-Q− complex

The primary processes of bacterial photosynthesis take place in a membrane-bound protein called the reaction center (RC). The process involves light-induced electron transfer from a primary electron donor to a sequence of electron acceptors. The terminal acceptors, operating sequentially, are two ubiquinones, Q_A and Q_B, that couple magnetically to a high-spin (S = 2) Fe²⁺ forming an Fe²⁺-Q^? complex. We have used a variety of techniques to investigate the electronic as well as some features of the spatial structure of the Fe²⁺-Q^? complex in RCs fromRb. sphaeroides. These include: (a) static magnetization measurements in the temperature range of 0.7 to 190 K and magnetic fields up to 8 kG, (b) EPR spectroscopy at helium temperatures at 1.2, 9, and 35 GHz, (c) Mössbauer spectroscopy and (d) extended X-ray fine structure (EXAFS) determinations. The results of these measurements showed that Fe²⁺ resides in an asymmetric ligand field environment forming 6 ligands with a combination of oxygens and nitrogens. From the EXAFS results the distances of the Fe²⁺ to the first and third coordination shells were determined. These spatial features were subsequently corroborated by the X-ray structure of the RC, which showed the environment of the Fe to be a distorted octahedron, the base plane of which is formed by three N_?’s of histidines and one carbonyl oxygen; the apex is formed by a fourth N_? and a second carbonyl oxygen. The distances from the Fe²⁺ to the first and third shell were in good agreement with the values obtained from EXAFS. The most detailed information of the electronic structure of the Fe²⁺-Q^? complex was obtained from the EPR spectra using the spin Hamiltonian formalism. The following spin-Hamiltonian parameters were obtained: for the crystalline field parameters,D = 7.60 K,E/D = 0.25; for the electronicg-values, g_{Fe, x} = 2.16, g_{Fe, y} = 2.27, g_{Fe, z} = 2.04 and for the antiferromagnetic exchange interaction,J _x = ?0.13 K,J _y = ?0.58 K,J _z = ?0.58 K. The use of the spin Hamiltonian was validated by comparing the results obtained from it with those obtained from an exact numerical solution of the 25 lowest energy levels of the orbital Hamiltonian. Possible roles that Fe²⁺ may play in the function of the RC are discussed. They include a structural role in which Fe²⁺ liganded to four histidine nitrogens imparts a rigid structure to a four α-helix bundle and a role in the electron transfer kinetics, most notably from the intermediate acceptor I^? (bacteriopheophytin) to Q_A. Replacement of Fe²⁺ by diamagnetic Zn²⁺ retained all observed native characteristics of the RC.     

Diffraction at HERA, Color Opacity and Nuclear Shadowing in DIS off nuclei

The QCD factorization theorem for diffractive processes in DIS is used to derive formulae for the leading twist contribution to the nuclear shadowing of parton distributions in the low thickness limit (due to the coherent projectile (photon) interactions with two nucleons). Based on the current analyzes of diffraction at HERA we find that the average strength of the interactions which govern diffraction in the gluon sector at x≤ 10⁻³, Q ₀= 2 GeV is ∼50mb. This is three times larger than in the quark sector and suggests that applicability of DGLAP approximation requires significantly larger Q ₀ in the gluon sector. We use this information on diffraction to estimate the higher order shadowing terms due to the photon interactions with N≥ 3 nucleons which are important for the scattering of heavy nuclei and to calculate nuclear shadowing and Q ² dependence of gluon densities. For the heavy nuclei the amount of the gluon shadowing: G _A(x,Q ₀ ²) /AG _N(x,Q ₀ ²)_{|x ≤ 10−3}∼ 0.25–0.4 is sensitive to the probability of the small size configurations within wave function of the gluon “partonometer” at the Q ₀ scale. At this scale for A∼ 200 the nonperturbative contribution to the gluon density is reduced by a factor of 4–5 at x≤ 10⁻³ unmasking PQCD physics in the gluon distribution of heavy nuclei. We point out that the shadowing of this magnitude would strongly modify the first stage of the heavy ion collisions at the LHC energies, and also would lead to large color opacity effects in eA collisions at HERA energies. In particular, the leading twist contribution to the cross section of the coherent J/ψ production off A≥ 12 nuclei at s ⁻²≥ 70 GeV is strongly reduced as compared to the naive color transparency expectations. The Gribov black body limit for F _2A(x,Q ²) is extended to the case of the gluon distributions in nuclei and shown to be relevant for the HERA kinematics of eA collisions. Properties of the final states are also briefly discussed. Received: 12 March 1999     

Point defects in FeAl

Summary Point defects in annealed B2-phase FeAl samples in the range 47–53 at.% Fe were studied using⁵⁷Fe M?ssbauer spectroscopy. Spectra were analyzed using local environment models according to which point defects in atomic shells close to probe atoms induce shifts in the nuclear monopole interaction. For well-annealed samples, better results were obtained assuming only the presence of Fe_Al antisite and V_Fe vacancy defects, and not of Al_Fe antisite defects. Monopole interactions of⁵⁷Fe probes on the Fe and Al sublattices having no defects in the first two shells were about +0.27 and −0.03 mm s⁻¹, respectively, with respect to Fe in alpha-Fe metal. The shifts induced by Fe_Al and V_Fe defects in the first shells of Fe probes on the Fe and Al sublattices were −0.15 and −0.24 mm s⁻¹, respectively, and, in the second shells, +0.06 and +0.011 mm s⁻¹. In addition to structural defects needed to accommodate deviations from stoichiometry, annealed samples were found to contain several percents of Fe_Al and V_Fe defects due to lattice disorder, with greater disorder in Fe-deficient alloys. Paper presented at the ICAME-95, Rimini, 10–16 September 1995.     

Comparison of electron spin polarization of P 870 + Qa a − observed in Zn2+-containing and Fe2+-depleted bacterial reaction centers

Recently, a general model has been developed to explain electron spin polarized (ESP) electron paramagnetic resonance (EPR) signals found in systems where radical pairs are formed sequentially. The photosynthetic bacterial reaction center (RC) is such a system in which we can experimentally vary parameters (lifetime, structure, and magnetic interactions in the sequentially formed radical pairs) that affect ESP development in order to test this model. In Fe²⁺-depleted transfer step from intermediate radical pair, P ₈₇₀ ⁺ Q _a ^? which is produced in an electron transfer step from intermediate radical pair, P ₈₇₀ ⁺ I^?. (P ₈₇₀ ⁺ is the oxidized primary donor, a special pair of bacteriochlorophyll molecules, I^? is the reduced bacteriopheophytin acceptor, and Q _a ^? is the reduced primary quinone acceptor.) The lifetime of P ₈₇₀ ⁺ I^? can be shortened relative to the lifetime of P ₈₇₀ ⁺ I^? in Fe²⁺-depleted RCs by substitution of Zn²⁺. We report the first observation of X-band and Q-band ESP EPR signals due to P ₈₇₀ ⁺ Q^? from bacterial reaction centers that contain Zn²⁺. Comparison of these signals to those observed from Fe²⁺-depleted bacterial reaction centers shows intensity differences and g-factor shifts. The results are discussed in terms of the general sequential radical pair model.     

Domain state–dependent magnetic formation of Fe<Subscript>3</Subscript>O<Subscript>4</Subscript> nanoparticles analyzed via magnetic resonance

Magnetic properties, arising from surface exchange and interparticle interactions of the Fe₃O₄ (magnetite) nanoparticles, were investigated in the temperature range of 5–300 and 120–300 K using vibrating sample magnetometer technique and electron spin resonance spectroscopy, respectively. The research was based on to figure out the origin of intraparticle interactions and the change of interparticle interactions in wide size range Fe₃O₄ nanoparticles. The analyses were done for samples having almost same particle size distributions. The average particle sizes were changed in between 30 ± 2 and 34 ± 2 nm. The observed magnetization values were demonstrated the mixture of single-domain size particles, exhibiting both single-domain (SD) and superparamagnetic (SPM) states. The symmetry of resonance curves changed according to the ratio of SD and SPM-stated particles in mixture under located temperature. The changes of anisotropy up to domain state were understood by freezing magnetic moment in glycerol matrix from room temperature to 120 K under 5-kG field. The shift of H _R values to higher magnetic fields and the more symmetric resonance spectrum proved the effect of anisotropy and interparticle interactions fields on magnetic behave. In addition, the origin of intra-interaction was exposed from Fe³⁺ centers and exchange coupling in between Fe²⁺, Fe³⁺, and O⁻, and Fe³⁺ centers found from g factor (g).     

EPR and optical spectroscopy of impurities in two synthetic beryls

Two synthetic beryls (Al₂Be₃Si₆O₁₈) of different color (purple and blue-green) were studied with electron paramagnetic resonance (EPR) and optical spectroscopy. In both crystals, the known spectra of Cu²⁺ and Fe³⁺ were observed with the same relative intensity. In the purple sample heated at 700°C in hydrogen atmosphere, two different kinds of Mn²⁺ EPR spectra were observed. The main one is pseudoaxial, it arises from ions substituted for Al³⁺ at position 4c of the structure. The weaker one is more complex, it has orthorhombic symmetry and is characterized by an unusually large zero-field splitting (B ₂⁰ = 741 · 10⁻⁴ cm⁻¹) and an isotropic hyperfine constantA = 70 G. This spectrum arises from Mn²⁺ at position 6f in the structure, normally occupied by Be. From optics, the blue-green color arises from Cu²⁺, while the purple one is due to Mn³⁺.     

Spectral properties of chlorines and electron transfer with their participation in the photosynthetic reaction center of photosystem II

Structural factors that provide localization of excited states and determine the properties of primary donor and acceptor of electron in the reaction center of photosystem II (PSII RC) are studied. The results of calculations using stationary and time-dependent density functional theory indicate an important role of protein environments of chlorophylls P_A, P_B, B_A, and B_B and pheophytins H_A and H_B in the area with a radius of no greater than ≤10 Å in the formation of excitonic states of PSII RC. When the neighboring elements are taken into account, the wavelength of long-wavelength Q _y transition of chlorophyll molecules is varied by about 10 nm. The effect is less developed for pheophytin molecules (Δλ ? 2 nm). The following elements strongly affect energy of the transition: His^A198 and His^D197 amino-acid residues that serve as ligands of magnesium atoms affect P_A and P_B, respectively; Met^A183 affects P_A; Met^A172 and Met^D198 affect B_A; water molecules that are located above the planes of the B_A and B_B macrocycles form H bonds with carbonyl groups; and phytol chains of P_A and P_B affect B_A, B_B, H_A, and H_B. The analysis of excitonic states, mutual positions of molecular orbitals of electron donors and acceptors, and matrix elements of electron transfer reaction shows that (i) charge separation between B_A and H_A and P_B and B_A is possible in the active A branch of cofactors of PSII RC and (ii) electron transfer is blocked at the B_B - H_B fragment in inactive B branch of PSII RC.     

Factors determining the energy yield of luminophors based on single-crystal films of Al2−Y2O3−R2O3 oxides

We consider the factors determining the energy yield of crystallophosphors based on single-crystal films of oxides of the system Al₂−Y₂O₃−R₂O₃ (R is the rare-earth ions) that are used as different types of luminescent converters. Special features of producing the films by the method of liquid-phase epitaxy from Pb−B₂O₃ and Bi₂O₃-based fluxes are analyzed, and the advantages of this method for production of efficient luminophors, which are based on high-melting oxides doped with mercury-like impurities, are shown. It is established that the main factor that bounds the crystallophosphor luminous yield is the presence of donor-acceptor complexes of the Pb²⁺−Pb⁴⁺ and Fe³⁺−Fe²⁺-type that form different channels of dissipation of excitation energy. The means of minimizing the contribution of these ions to the processes of excitation-energy relaxation are discussed. Institute of Applied Physics, I. Franko L'vov State University, 49 General Chuprynka Str., L'vov, 290044, Ukraine. Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 66, No. 6, pp. 819–823, November–December, 1999.     

Mössbauer study of the spinel system GexCu1−xFe2O4

The magnetic properties of the spinel series Ge_xCu_1?xFe₂O₄ (X = 0 to 0.8) have been investigated by means of Mössbauer spectroscopy. Mössbauer spectra for X = 0.0 to 0.6 suggest the existence of two hyperfine fields, one due to the Fe³⁺ tetrahedral ions (A-sites) and the other due to Fe³⁺ octahedral ions (B-sites), while for X = 0.8 it shows the superposition of hyperfine field split spectra from A- and B-site ions and a broad central line spectrum. For 0.2 ? X ? 0.4, fast electron exchange among octahedral iron ions occurs as in Fe₃O₄. The variations of nuclear magnetic fields at the A- and B-sites are explained on the basis of AB and BB supertransferred hyperfine interactions.     

Semi-quantitative theory of the structures of simple ionic crystals

A simple theory is developed which shows that the regions of stability of the CsCl, NaCl and ZnS structures can be demarcated in a two-dimensional plot of the radius ratio versus the strength of the van der Waals interaction. There is good agreement with experiment. The effect of pressure on these structures is explained qualitatively. The increased occurrence of the ZnS structure and the decreased stability of the CsCl structure in the A²⁺ B²⁻ crystals compared to the A⁺B⁻ crystals is also explained. Finally it is shown that the radius ratio and the polarizabilities of the ions are the important factors that determine the structures of AB₂ crystals.     

Oxygen-dependent Oxidation of Fe(II) to Fe(III) and Interaction of Fe(III) with Bovine Serum Albumin, Leading to a Hysteretic Effect on the Fluorescence of Bovine Serum Albumin

The serum albumin is the most abundant protein in blood plasma and the iron is essential for many cellular processes. However, the interaction between Fe³⁺ and haem-free serum albumin remains unclear. Here we provide evidence for the fact that haem-free BSA possesses one specific Fe³⁺-binding site. The binding of Fe³⁺ to BSA results in a significant quenching of the Trp fluorescence of BSA. The average apparent dissociation constant value for the interaction of Fe³⁺ and BSA is 3.46 × 10⁻⁸ ± 3 × 10⁻¹⁰ M at 37 °C and 3.30 × 10⁻⁸ ± 5 × 10⁻¹⁰ M at 25 °C, respectively, as determined by fluorescence titration. Addition of 50 μM Fe²⁺ to 1 μM BSA results in an obvious hysteretic effect on the fluorescence of BSA. The time-dependent fluorescence quenching of BSA by Fe²⁺ is not caused by the Fe²⁺-induced conformational change of BSA, but the oxygen-dependent oxidation of Fe²⁺ to Fe³⁺. Fe²⁺ undergoes an oxygen-dependent oxidation to Fe³⁺ under aerobic conditions, which is accelerated by the interaction of BSA with Fe³⁺ and extensively inhibited under anaerobic conditions. The results suggest that BSA may take part in non-transferrin bound iron transfer.