Preparation protocols for high-activity photosystem II membrane particles of green algae and higher plants, pH dependence of oxygen evolution and comparison of the S2-state multiline signal by X-band EPR spectroscopy

Photosystem II (PS II) membrane particles are particularly well suited for various types of spectroscopic investigations on the PS II manganese complex. Here we present: (1) a preparation protocol for PS II membrane particles of higher plants, which yields exceptionally high oxygen-evolution activity due to the use of glycinebetaine as a PS II-stabilizing agent; (2) preparation protocols for highly active PS II membrane particles for the green algae Scenedesmus obliquus and Chlamydomonas reinhardtii; (3) a determination of pH dependence of oxygen evolution for spinach and Scenedesmus; (4) a comparison of the EPR multiline signal observed in the S2-state of green algae and higher plants of PS II membrane particles. A clearly broader type of multiline EPR signal is observed in green algae.     

g-Anisotropy of the S2-state manganese cluster in single crystals of cyanobacterial photosystem II studied by W-band electron paramagnetic resonance spectroscopy

The multiline signal from the S2-state manganese cluster in the oxygen evolving complex of photosystem II (PSII) was observed in single crystals of a thermophilic cyanobacterium Thermosynechococcus vulcanus for the first time by W-band (94 GHz) electron paramagnetic resonance (EPR). At W-band, spectra were characterized by the g-anisotropy, which enabled the precise determination of the tensor. Distinct hyperfine splittings (hfs's) as seen in frozen solutions of PSII at X-band (9.5 GHz) were detected in most of the crystal orientations relative to the magnetic field. In some orientations, however, the hfs's disappeared due to overlapping of a large number of EPR lines from eight crystallographic symmetry-related sites of the manganese cluster within the unit cell of the crystal. Analysis of the orientation-dependent spectral features yielded the following g-tensor components: g(x) = 1.988, g(y) = 1.981, g(z) = 1.965. The principal values suggested an approximate axial symmetry around the Mn(III) ion in the cluster.     

EPR and optical spectroscopic studies of Met80X mutants of yeast ferricytochrome c. Models for intermediates in the alkaline transition

The ferric forms of Met80X mutants of yeast iso-1-cytochrome c (X = Ala, Ser, Asp, and Glu) display EPR and optical spectra that are strongly pH dependent. At low pH values (pH approximately 5) the sixth coordination sites are filled by H(2)O that, on elevating the pH, is replaced by OH(-) in the cases of Met80Ala and -Ser (pK approximately 5.6 and 5.9, respectively) and by a lysine amino group in the cases of Met80Asp and -Glu (pK approximately 9.3 and 11.6, respectively). The ligand sets and the pK values of the transitions are rationalized in terms of the structure of the heme pocket, and a possible mechanism of the "trigger" in the alkaline transition of the native protein is suggested.     

Differential effects of plasma membrane electric excitation on H+ fluxes and photosynthesis in characean cells

Cells of characean algae exposed to illumination arrange plasma-membrane H(+) fluxes and photosynthesis in coordinated spatial patterns (bands). This study reveals that H(+) transport and photosynthesis patterns in these excitable cells are affected not only by light conditions but also by electric excitation of the plasma membrane. It is shown that generation of action potential (AP) temporally eliminates alkaline bands, suppresses O(2) evolution, and differentially affects primary reactions of photosystem II (PSII) in different cell regions. The quantum yield of PSII electron transport decreased after AP in the alkaline but not in acidic cell regions. The effects of electric excitation on fluorescence and the PSII electron flow were most pronounced at light-limiting conditions. Evidence was obtained that the shift in chlorophyll fluorescence after AP is due to the increase in DeltapH at thylakoid membranes. It is concluded that the AP-triggered pathways affecting ion transport and photosynthetic energy conversion are linked but not identical.     

Heme-Peptide Models for Hemoproteins. 1. Solution Chemistry of N-Acetylmicroperoxidase-8

An improved method for the preparation of the heme octapeptide acetyl-MP8, obtained by proteolysis of horse heart cytochrome c, is described. AcMP8 obeys Beer's law at pH 7.0 in aqueous solution up to a concentration of 3 x 10(-)(5) M. The self-association constant measured at 25 degrees C (log K(D) = 4.04) is an order of magnitude lower than that for MP8, reflecting the role of the N-acetyl protecting group in abolishing intermolecular coordination. However, AcMP8 does form pi-stacked dimers in aqueous solution with increasing ionic strength. A more weakly packed pi-pi dimer reaches a maximum abundance at approximately 3 M ionic strength, but a more tightly packed dimer is favored at &mgr; > 3 M. An equilibrium model based on charge neutralization by specific binding of Na(+) ions gives a total molecular charge of 3- for AcMP8 at pH 7.0 and a self-association constant log K(D) = 4.20. AcMP8 exhibits six spectroscopically active pH-dependent transitions. The Glu-21 c-terminal carboxylate binds to the heme iron at low pH (pK(a) = 2.1) but is substituted by His-18 (pK(a) = 3.12) as the pH increases. The two heme propanoic acid substituents ionize with pK(a)'s of 4.95 and 6.1. This is followed by ionization of iron-bound water with a pK(a) = 9.59, DeltaH = 48 +/- 1 kJ mol(-)(1), and DeltaS = -22 +/- 3 J K(-)(1) mol(-)(1). The electronic spectra indicate that AcMP8 is predominantly in the S = (5)/(2) state at pH 7.0, while the hydroxo complex at pH 10.5 corresponds to an equilibrium mixture of S = (5)/(2) and S = (1)/(2) states at 25 degrees C. In the final transition, His-18 ionizes to form the S = (1)/(2) histidinate complex with a pK(a) of 12.71. AcMP8 is relatively stable under alkaline conditions, dimerizing slowly at high pH (k = 2.59 +/- 0.14 M(-)(1) s(-)(1)) to form a high-spin &mgr;-oxo-bridged species. The pH-dependent behavior of AcMP8 in the presence of excess 3-cyanopyridine, however, is markedly different. At low pH, AcMP8 simultaneously binds the exogenous ligand and the Glu-21 c-terminal carboxylate with a pK(a) < 2. His-18 replaces the carboxylate ligand at higher pH (pK(a) = 2.60), and both heme propanoic acid groups ionize with a mean pK(a) = 5.10. Unlike AcMP8.OH(-), the axial histidine of the 3-CNPy complex ionizes at near neutral pH (pK(a) = 7.83), prior to being replaced by OH(-) (pK(a) = 10.13). The sixth transition in the AcMP8/3-CNPy system produces the bis(hydroxo) complex (pK(a) > 13).     

Thermodynamic, kinetic and pH studies on the reactions of NCS-, N3-, and CH3CO2- with Fusarium galactose oxidase

Thermodynamic and kinetic studies on the X- = NCS-, N3-, and CH3CO2- replacement of H2O/OH- at the CuII exogenous site of the tyrosyl-radical-containing enzyme galactose oxidase (GOaseox) from Fusarium (NRR 2903), have been studied by methods involving UV-vis spectrophotometry (25 degrees C), pH range 5.5-8.7, I = 0.100 M (NaCl). In the case of N3- and CH3CO2- previous X-ray structures have confirmed coordination at the exogenous H2O/OH- site. From the effect of pH on the UV-vis spectrum of GOaseox under buffer-free conditions, acid dissociation constants of 5.7 (pK1a; coordinated H2O) and 7.0 (pK2a; H+Tyr-495) have been determined. At pH 7.0 formation constants K(25 degrees C)/M-1 are NCS- (480), N3- (1.98 x 10(4)), and CH3CO2- (104), and from the variations in K with pH the same two pKa values are seen to apply. No pK1a is observed when X- is coordinated. From equilibration stopped-flow studies rate constants at pH 7.0 for the formation reaction kf(25 degrees C)/M-1 s-1 are NCS- (1.13 x 10(4)) and N3- (5.2 x 10(5)). Both K and kf decrease with increasing pH, consistent with the electrostatic effect of replacing H2O by OH-. In the case of the GOaseox Tyr495Phe variant pK1a is again 5.7, but no pK2a is observed, confirming the latter as acid dissociation of protonated Tyr-495. At pH 7.0, K for the reaction of four-coordinate GOaseox Tyr495Phe with NCS- (1.02 x 10(5) M-1) is more favorable than the value for GOaseox. Effects of H+Tyr-495 deprotonation on K are smaller than those for the H2O/OH- change. The pK1a for GOasesemi is very similar (5.6) to that for GOaseox (both at CuII), but pK2a is 8.0. At pH 7.0 values of K for GOasesemi are NCS- (270 M-1), N3- (4.9 x 10(3)), and CH3CO2- (107).     

Lanthanide macrocyclic quinolyl conjugates as luminescent molecular-level devices.

The Eu(III) tetraazamacrocyclic complexes [Eu.1] and [Eu.2], and the Tb(III) and Yb(III) complexes [Tb.1] and [Yb.2], have been synthesized as luminescent molecular-level devices. The Eu complexes exhibit unique dual pH switching behavior in water under ambient conditions. The delayed Eu emission is reversibly switched on in acid, with an enhancement factor of several hundred for [Eu.1]. These observations are consistent with the protonation of the quinoline aryl nitrogen moiety (pK(a) approximately equal to 5.9 for [Eu.1]). The fluorescence emission spectra of these complexes are unaffected by acid, but pronounced changes occur in alkaline solution due to the deprotonation of the aryl amide nitrogen (pK(a) approximately 9.4 for [Eu.1]). [Tb.1] shows a more intriguing pH dependence; Tb emission is switched "on" only in the presence of H+ and in the absence of molecular oxygen, whereas the fluorescence emission properties are similar to those observed with [Eu.1]. This behavior can be conveniently described as a molecular-level logic gate, corresponding to a two-input INHIBIT function, A wedge B'. The analogous [Yb.2] complex shows no such pH or O(2) dependence.     

Relationship between the structural and functional changes of the photosynthetic apparatus during chloroplast-chromoplast transition in flower bud of Lilium longiflorum

The relationship between the structural and functional changes of the photosynthetic apparatus in the flower bud of Lilium longiflorum during chloroplast-chromoplast transition was examined. Compared with green petals, there was a five-fold increase of the carotenoid content in yellow petals, whereas the chlorophyll content decreased five-fold. Absorption and emission fluorescence spectra of chromoplasts indicated that newly synthesized carotenoids were not associated with photosystem II (PSII) photochemistry. The maximum quantum yield in the remaining PSII reaction centers remained constant during the chromoplast formation, whereas the photosynthetic electron transport beyond PSII became inhibited, as indicated by a marked decrease of the O2 evolution capacity, of the photochemical quenching of chlorophyll-alpha fluorescence and of the operational quantum yield of photosynthetic electron transport. Deconvoluted fluorescence emission spectra indicated a more rapid degradation of photosystem I (PSI) complexes than of PSII during chromoplast formation. Compared with green petals, the spillover between PSII and PSI decreased by approximately 40% in yellow petals. Our results indicate that during chloroplast-chromoplast transition in the flower bud of L. longiflorum, PSII integrity was preserved longer than the rest of the photosynthetic apparatus.     

Investigation of the calcium-binding site of the oxygen evolving complex of photosystem II using 87Sr ESEEM spectroscopy

The proximity of the calcium/strontium binding site of the oxygen evolving complex (OEC) of photosystem II (PSII) to the paramagnetic Mn cluster is explored with (87)Sr three-pulse electron spin-echo envelope modulation (ESEEM) spectroscopy. CW-EPR spectra of Sr(2+)-substituted Ca(2+)-depleted PSII membranes show the modified g = 2 multiline EPR signal as previously reported. We performed three-pulse ESEEM on this modified multiline signal of the Mn cluster using natural abundance Sr and (87)Sr, respectively. Three-pulse ESEEM of the natural abundance Sr sample exhibits no detectable modulation by the 7% abundance (87)Sr. On the other hand, that of the (87)Sr enriched (93%) sample clearly reveals modulation arising from the I = (9)/(2) (87)Sr nucleus weakly magnetically coupled to the Mn cluster. Using a simple point dipole approximation for the electron spin, analysis of the (87)Sr ESEEM modulation depth via an analytic expression suggests a Mn-Ca (Sr) distance of 4.5 A. Simulation of three-pulse ESEEM with a numerical matrix diagonalization procedure gave good agreement with this analytical result. A more appropriate tetranuclear magnetic/structural model for the Mn cluster converts the 4.5 A point dipole distance to a 3.8-5.0 A range of distances. DFT calculations of (43)Ca and (87)Sr quadrupolar interactions on Ca (and Sr substituted) binding sites in various proteins suggest that the lack of the nuclear quadrupole induced splitting in the ESEEM spectrum of (87)Sr enriched PSII samples is related to a very high degree of symmetry of the ligands surrounding the Sr(2+) ion in the substituted Ca site. Numerical simulations show that moderate (87)Sr quadrupolar couplings decrease the envelope modulation relative to the zero quadrupole case, and therefore we consider that the 3.8-5.0 A range obtained without quadrupolar coupling included in the simulation represents an upper limit to the actual manganese-calcium distance. This (87)Sr pulsed EPR spectroscopy provides independent direct evidence that the calcium/strontium binding site is close to the Mn cluster in the OEC of PSII.     

Shape-shifting tetranuclear oxo-bridged manganese cluster: relevance to photosystem II water oxidase active site

The redox properties of the "dimer-of-dimers" complex, [{Mn2(mu-O)2(tphpn)}2]4+ (1) (where Htphpn = N,N,N',N'-tetra(2-methylpyridyl)-2-hydroxypropane-diamine) were investigated. The structure changes dramatically to an adamantane-shaped core upon one-electron oxidation. On the other hand, the one-electron reduced product of 1, [Mn4O4(tphpn)2]3+, exhibits a hyperfine-structured multiline EPR signal very similar to the so-called S0 state of the tetramanganese cluster, which resides at the Photosystem II water oxidase active site.     

Mechanism of reaction of hydrogen peroxide with horseradish peroxidase: identification of intermediates in the catalytic cycle.

The mechanism of the reaction of horseradish peroxidase isoenzyme C (HRPC) with hydrogen peroxide to form the reactive enzyme intermediate compound I has been studied using electronic absorbance, rapid-scan stopped-flow, and electron paramagnetic resonance (EPR) spectroscopies at both acid and basic pH. The roles of the active site residues His42 and Arg38 in controlling heterolytic cleavage of the H(2)O(2) oxygen-oxygen bond have been probed with site-directed mutant enzymes His42 --> Leu (H42L), Arg38 --> Leu (R38L), and Arg38 --> Gly (R38G). The biphasic reaction kinetics of H42L with H(2)O(2) suggested the presence of an intermediate species and, at acid pH, a reversible second step, probably due to a neutral enzyme-H(2)O(2) complex and the ferric-peroxoanion-containing compound 0. EPR also indicated the formation of a protein radical situated more than approximately 10 A from the heme iron. The stoichiometry of the reaction of the H42L/H(2)O(2) reaction product and 2,2'-azinobis(3-ethylbenzothiazolinesulfonic acid) (ABTS) was concentration dependent and fell from a value of 2 to 1 above 0.7 mM ABTS. These data can be explained if H(2)O(2) undergoes homolytic cleavage in H42L. The apparent rate of compound I formation by H42L, while low, was pH independent in contrast to wild-type HRPC where the rate falls at acid pH, indicating the involvement of an ionizable group with pK(a) approximately 4. In R38L and R38G, the apparent pK(a) was shifted to approximately 8 but there is no evidence that homolytic cleavage of H(2)O(2) occurs. These data suggest that His42 acts initially as a proton acceptor (base catalyst) and then as a donor (acid catalyst) at neutral pH and predict the observed slower rate and lower efficiency of heterolytic cleavage observed at acid pH. Arg38 is influential in lowering the pK(a) of His42 and additionally in aligning H(2)O(2) in the active site, but it does not play a direct role in proton transfer.     

Transformation of the Photoacoustic Signal after Treatment of Barley Leaves with Methylviologen or High Temperatures

Abstract— The photoacoustic (PA) signal at the modulation frequency of 35 Hz was studied in MV-treated barley leaves or leaves preheated at different temperatures. Saturating illumination enhanced the magnitude of the PA signal in MV-treated leaves in contrast with the opposite result usually found in untreated intact leaves where saturating illumination abolishes the photobaric component of the PA signal due to oxygen evolution and thus decreases the total PA signal. A linear relationship was found between the changes induced by continuous background light in the negative response of PA signal to saturating light in intact leaves and in the positive response in MV-treated leaves. A linear relationship was also observed in MV-treated leaves between the positive changes in the PA signal and the changes in the rate of electron transport through photosystem II (PSII) calculated from chlorophyll fluorescence data. The conclusion was drawn that only the thermal component contributes to the PA signal measured at low modulated frequency in MV-treated leaves because the enhanced O2 uptake provides a zero net oxygen exchange by superimposing with O2 evolution. The leaves preheated at temperatures above 43°C demonstrated the positive response of the PA signal to saturating light at 35 Hz. In leaves preheated at 41.5°C, the first and second saturating pulses induced the enhancement of PA signal, whereas other pulses decreased the PA signal due to onset of oxygen evolution. The energy storage activity measured in the absence of oxygen evolution in heat-treated leaves is proposed to be associated with cyclic electron transport activities around PSII and PSI.     

An electron paramagnetic resonance study of Mn2(H2O)(OAc)4(tmeda)2 (tmeda = N,N,N',N'-tetramethylethylenediamine): a model for dinuclear manganese enzyme active sites

The complex Mn2(H2O)(OAc)4(tmeda)2 (tmeda = N,N,N',N'-tetramethylethylenediamine) is a model for the active site of hydrolase enzymes containing acetate-bridged dimanganese cores. The two high-spin Mn(II) ions are antiferromagnetically coupled, as determined by previous magnetic susceptibility studies (Yu, S.-B; Lippard, S. J.; Shweky, I; Bino, A. Inorg. Chem. 1992, 31, 3502-3504) to yield a spin "ladder" with total spin S = 0, 1, 2, ..., 5 in increasing energy. In this study, the complex was characterized by Q-band and X-band EPR spectroscopy in frozen solution. Analysis of the temperature dependence of these EPR spectra indicates that the primary spectral contribution is from the S = 2 manifold. The EPR spectra were simulated using a full spin Hamiltonian for this manifold of a coupled spin system, which provided the fit parameters J = -2.9 cm-1, g = 2.00, and D2 = -0.060 +/- 0.003 cm-1. An additional multiline EPR signal is observed which is proposed to arise from the total spin S = 5/2 ground state of a Mn(II) trimer of the type Mn3(OAc)6(tmeda)2.     

On the relation between the Kautsky effect (chlorophyll a fluorescence induction) and Photosystem II: basics and applications of the OJIP fluorescence transient

Chlorophyll a fluorescence is a highly sensitive, non-destructive, and reliable tool for measuring, rather quickly, photosynthetic efficiency, particularly of Photosystem II (PSII), the water-plastoquinone oxidoreductase. We briefly review here the connection between the fast (up to 2 s) chlorophyll fluorescence rise and PSII, as well as the empirical use of the fluorescence rise kinetics in understanding photosynthetic reactions, particularly of PSII. When dark-adapted photosynthetic samples are exposed to light, a fluorescence induction is observed, known as the Kautsky effect, after Hans Kautsky, the discoverer of the phenomenon showing the existence of variable fluorescence. The chlorophyll fluorescence intensity rises from a minimum level (the O level), in less than 1 s, to a maximum level (the P-level) via two intermediate steps labeled J and I. This is followed by a decline to a lower semi-steady state level, the S level, which is reached in about one minute. We provide here an educational review on how this phenomenon has been exploited through analysis of the fast OJIP fluorescence transient, by discussing basic assumptions, derivation of equations, as well as application to PSII-related questions.     

The effect of pyridinecarboxylate chelating groups on the stability and electronic relaxation of gadolinium complexes

The ligand N,N'-bis[(6-carboxy-2-pyridylmethyl]ethylenediamine-N,N'-diacetic acid (H(4)bpeda) was synthesised using an improved procedure which requires a reduced number of steps and leads to a higher yield with respect to the published procedure. It was obtained in three steps from diethylpyridine-2,6-dicarboxylate and commercially available ethylenediamine-N,N[prime or minute]-diacetic acid with a total yield of approximately 20%. The crystal structure of the hexa-protonated form of the ligand which was determined by X-ray diffraction shows that the four carboxylates and the two amines are protonated. The crystal structure of the polynuclear complex [Gd(bpeda)(H(2)O)(2)](3)[Gd(H(2)O)(6)](2)Cl(3)(2), isolated by slow evaporation of a 1:1 mixture of GdCl(3) and H(4)bpeda at pH approximately 1, was determined by X-ray diffraction. In complex three [Gd(bpeda)(H(2)O)(2)] units, containing a Gd(III) ion ten-coordinated by the octadentate bpeda and two water molecules, are connected in a pentametallic structure by two hexa-aquo Gd(3+) cations through four carboxylato bridges. The protonation constants (pK(a1)= 2.9(1), pK(a2)= 3.5(1), pK(a3)= 5.2(2), and pK(a4)= 8.5(1)) and the stability constants of the complexes formed between Gd(III) and Ca(II) ions and H(4)bpeda (log beta(GdL)= 15.1(3); log beta(CaL)= 9.4(1)) were determined by potentiometric titration. The unexpected decrease in the stability of the gadolinium complex and of the calcium complex of the octadentate ligand bpeda(4-) with respect to the hexadentate ligand edta(4-) has been interpreted in terms of an overall lower contribution to stability of the metal-nitrogen interactions. The EPR spectra display very broad lines (apparent DeltaH(pp) approximately 800-1200 G at X-band and 90-110 G at Q-band depending on the temperature), indicating a rapid transverse electron spin relaxation. At X-band, Gd(bpeda) is among the fastest relaxing Gd(3+) complexes to date suggesting that the presence of pyridinecarboxylate chelating groups in itself does not lead to slow electron relaxation.     

Polarography of uranium(VI) and lead(II) complexes with l-glutamine

The complexes of uranium(VI) and lead(II) with 1-glutamine were investigated polarographically. For uranium(VI), the complexes UO(2)G(+2), UO(2)G(2)(+2) and UO(2)(OH)Ga(2)(+) were identified at pH < 2.5, pH 2.5-4.1 and pH 4.1-5.2 respectively. With lead(II), complexes PbG(+2), Pb(OH)G(+) and Pb(OH)G(2)(+) were formed at pH 2.0-5.0, pH 5.0-7.0, and pH 7.0-8.5, respectively. The concentration dissociation constant of Pb(OH)G(2)(+) was found to be pK(c) = 10.16 +/- 0.04 at ionic strength 0.6.     

Synthesis of thiomorpholino buffers for isoelectric focusing in immobilized pH gradients

The two commercially available Immobilines having a pK of 6.2 (2-morpholino ethyl acrylamide) and 7.0 (3-morpholinopropylacrylamide) have been modified and two new buffers have been synthesized: 2-thiomorpholinoethylacrylamide, pK 6.6, and 3-thiomorpholinopropyl acrylamide, pK 7.4. The replacement of an oxygen with a sulfur atom in the morpholino ring is thus seen to shift the pK values of these two bases by +0.4 pH units. In formulations in which the two new bases replaced the standard morpholino derivatives, identical pH profiles and protein patterns were obtained. The reason for this work was to try to close the gap between the pK 7.0 and 8.5 species and to provide the users of immobilized pH gradients with more buffers in the neutral pH region. The two new thiomorpholino derivatives are an important step in this direction.     

The peroxidase activity of a hemin--DNA oligonucleotide complex: free radical damage to specific guanine bases of the DNA.

A specific DNA oligonucleotide--hemin complex (PS2.M--hemin complex) that exhibits DNA-enhanced peroxidative activity was studied by EPR and UV--visible spectroscopy and by chemical probing analysis. EPR data obtained from low-temperature experiments on the PS2.M--hemin complex showed both a low-field g approximately 6 and a high-field g approximately 2 signal. These EPR signals are typical of high-spin ferric heme with axial symmetry as judged by the EPR spectrum of six-coordinate heme iron in acidic Fe(III)-myoglobin. This similarity is consistent with the presence of two axial ligands to the heme iron within the PS2.M--hemin complex, one of which is a water molecule. Optical analyses of the acid-base transition for the hemin complex yielded a pK(a) value for the water ligand of 8.70 +/- 0.03 (mean +/- SD). Low-temperature EPR analysis coupled with parallel spin-trapping investigations following the reaction of the PS2.M--hemin complex and hydrogen peroxide (H(2)O(2)) indicated the formation of a carbon-centered radical, most likely on the PS2.M oligonucleotide. Chemical probing analysis identified specific guanine bases within the PS2.M sequence that underwent oxidative damage upon reaction with H(2)O(2). These and other experimental findings support the hypothesis that the interaction of specific guanines of PS2.M with the bound hemin cofactor might contribute to the superior peroxidative activity of the PS2.M--hemin complex.     

Decomposition kinetics of Ni(III)-peptide complexes with histidine and histamine as the third residue

The decomposition kinetics of the Ni(III) complexes of Gly(2)HisGly and Gly(2)Ha are studied from p[H(+)] 3.5 to 10, where His is l-histidine and Ha is histamine. In these redox reactions, at least two Ni(III) complexes are reduced to Ni(II) while oxidizing a single peptide ligand. The rate of Ni(III) loss is first order at low pH, mixed order from pH 7.0 to 8.5, and second order at higher pH. The transition from first- to second-order kinetics is attributed to the formation of an oxo-bridged Ni(III)-peptide dimer. The rates of decay of the Ni(III) complexes are general-base assisted with Br?nsted beta values of 0.62 and 0.59 for Ni(III)Gly(2)HisGly and Ni(III)Gly(2)Ha, respectively. The coordination of Gly(2)HisGly and Gly(2)Ha to Ni(II) are examined by UV-vis and CD spectroscopy. The square planar Ni(II)(H(-2)Gly(2)HisGly)(-) and Ni(II)(H(-2)Gly(2)Ha) complexes lose an additional proton from an imidazole nitrogen at high pH with pK(a) values of 11.74 and 11.54, respectively. The corresponding Ni(III) complexes have axially coordinated water molecules with pK(a) values of 9.37 and 9.44. At higher pH an additional proton is lost from the imidazole nitrogen with a pK(a) value of 10.50 to give Ni(III)(H(-3)Gly(2)Ha)(H(2)O)(OH)(2-).     

Fiber optic pH sensing with long wavelength excitable Schiff bases in the pH range of 7.0-12.0

Most of the fluorescent pH probes work near neutral or acidic regions of the pH scale. In this work, two different fluorescent Schiff bases, chloro phenyl imino propenyl aniline (CPIPA) and nitro phenyl imino propenyl aniline (NPIPA), have been investigated for pH sensing in the alkaline region. Absorption and emission based spectral data, quantum yield, fluorescence lifetime, photostability and acidity constant (pK(a)) of the Schiff bases were determined in conventional solvents and in PVC. The long wavelength excitable immobilized Schiff bases CPIPA (lambda(ex)=556 nm) and NPIPA (lambda(ex)=570 nm) exhibited absorption and emission based optical response to proton in the pH range of 8.0-12.0 and 7.0-12.0, respectively. Response of the CPIPA was fully reversible within the dynamic working range. The response times were between 3-13 min. A relative signal change of 95% and 96% have been achieved for sensor dyes of CPIPA and NPIPA, respectively. The CPIPA displayed better fluorescence quantum yield (varphi(F)=3.7 x 10(-1)) and higher matrix compatibility compared to NPIPA (varphi(F)=1.6 x 10(-1)) in immobilized PVC. The CPIPA and NPIPA exhibited a slight cross sensitivity to the ions of Hg(+) and Fe(3+), respectively.