THE KINETICS OF THE EARLY STAGES OF THE PHYTOCHROME PHOTOTRANSFORMATION Pr→ Pfr. A COMPARATIVE STUDY OF SMALL (60 kDalton) and NATIVE (124 kDalton) PHYTOCHROMES FROM OAT

A comparative study of the decay kinetics of photogenerated transients from small (60 kDalton) and native (124 kDalton) oat phytochrome in the red-absorbing form (P_r) in phosphate buffer containing 5 mM ethylenediamine tetraacetic acid, pH 7.8, (PB) and in PB containing 20% ethylene glycol, has been carried out in the temperature range 275–298 K. The analysis confirmed that at least two primary photoproducts, intermediates Iⁱ₇₀₀s and Iⁱ_7oo are formed from P_r. The kinetic parameters, as observed in PB at 695 nm and 275 K, are similar for the I₇₀₀ intermediates of both small and native phytochrome. Namely, the lifetimes are about 21 μs (component percentages 38%) for the I Iⁱ₇₀₀s and about 200 μ.s (62%) for the Iⁱ₇₀₀S- Arrhenius preexponential factors (A) of about 10¹⁶ and 10¹⁵ s-¹and activation energies of about 61 and 56 kJ/mol were measured for the absorbance decays of the I₇₀₀S of small and native phytochrome, respectively. The kinetic data favour parallel paths for the formation of the Iⁱ₇₀₀s from P_r, and the activation parameters indicate that the primary photoreactions of the transformation from P_r to the far-red-absorbing form are restricted to the chromophore within the protein. Moreover, the relatively modest temperature dependence of the lifetimes of the Iⁱ₇₀₀^S from small and native P_r supports the working hypothesis that the ground state reactions to the Iⁱ_bl, intermediates–although somewhat influenced by the polypeptide fragment that is removed upon degradation of native to small P_r–are localized to the chromophore, as is most probably the case also for the primary photoreactions. The effect of the addition of 20% ethylene glycol on the pre-exponential factors of the time-dependent decay functions is discussed in similar terms of the early stages of the phototransformation.     

INTERMEDIATES IN THE PHOTOCONVERSION OF FUNCTIONAL PHYTOCHROME IN FERN SPORES OF Dryopteris–II. In vivo KINETICS OF THE DECAY OF Ii700 AND OF THE FORMATION OF Pfr STUDIED WITH A DOUBLE-LASER APPARATUS*

Abstract— Photoconversion of the red-light-absorbing form of phytochrome, P_r, to the far-red-light-absorbing form, P_fr, was investigated in vivo at 22°C with 600 or 800 ns laser pulses of high spectral purity and induction of spore germination in Dryopteris paleacea was used as indicator for the progress of photoconversion. This reaction is initiated by a saturating R-laser pulse of 648.5 nm, establishing an equilibrium of the photochromic system between P_r and the very early intermediates, Iⁱ₇₀₀ (P_rφ Iⁱ₇₀₀)- The decay of Iⁱ₇₀₀ as well as the formation of P_fr was recorded by the application of a second pulse varied between 698 and 717.5 nm, which inhibits the formation of P_lr being absorbed predominantly by Iⁱ₇₀₀or P_fr, respectively. The most effective inhibition for the second pulse is found up to 10 u.s after the first pulse and this is interpreted by photoreversion of Iⁱ₇₀₀ to P_r; thus reducing the formation of P_fr from Iⁱ₇₀₀. This early inhibition decreases between 10 μs to 100 ms after the R-laser pulse, as a result of the decay of Iⁱ_bl to a bleached species I,;. This decay can be described by three first order kinetics with the rate constants k₁₂= 16830 ± 2970 s^-1, k₁₂= 666 ± 218 s^-1,k₁₃= 9.8 ± 0.9 s^-1. A second inhibition, due to the formation of P_fr, is found for dark intervals <100 ms and can be described by two first order kinetics with the rate constants k₂₁= 2.9 ± 0.6 s^-1 and k₂₂= 0.17 s^-l.     

INTERMEDIATES IN THE PHOTOCONVERSION OF FUNCTIONAL PHYTOCHROME IN FERN SPORES OF Dryopteris-I DEMONSTRATION AND QUANTITATIVE CHARACTERIZATION OF THE PHOTOCHROMIC SYSTEM Pr⇔ Ii700USING NANOSECOND-LASER PULSES*,†

Abstract— Spores of Dryopteris paleacea and D. filix-mas are positively photoblastic with an optimum in the action spectrum around 665 nm. Light is perceived by phytochrome and the relationship between germination and mole fraction of the far-red-absorbing form of this pigment, P_fr, was investigated with saturating irradiations between 662 and 747 nm under low-fluence-rate conditions. These control irradiations establish a proportion of the total phytochrome, P,_tot, as P_fr with P_fr/P_tot–φ at equilibrium. These φ -values were calculated according to data for native oat phytochrome (Kelly and Lagarias, 1985, Biochemistry 24, 6003) and the spectral characteristics of the interference filters. With this method a linear relationship could be found between φ and germination from 2 to 70% for D. paleacea and from 2 to 90% for D. filix-mas, if probit germination was plotted vs probit φ This correlation formed the basis of investigating the phytochrome photoconversion by dye-laser pulses of 380 ± 30 ns under high-fluence-rate conditions, and thus to test quantitatively the impact of the photoreversibility of intermediate reactions of the photoconversion and the red-absorbing form of phytochrome, P_fr on the final P_fr-level. Spore germination was initiated by a single-laser pulse in the range from 592 to 700 nm. The most effective wavelengths were 649 and 660 nm in both species, and at saturation maximal germination (ca. 50%) was obtained from 592 to 665 nm for D. paleacea or ca. 60% germination from 592 to 670 nm for D. filix-mas. Both saturation levels correspond to a ø-value between 0.40 and 0.45. This significantly diminished photoconversion is a consequence of the high-fluence-rate conditions during the laser pulse which establishes the photochromic system between P_r and a set of very early intermediates, Iⁱ_700, (= P_r? Iⁱ₇₀₀). This system can be described by the extinction coefficients of P_r and the intermediates Iⁱ₇₀₀, and by the quantum yields, 4,φ for the forward and reverse reactions as φ If φ is calculated, assuming a quantum yield of 1:1 for both reactions and with the extinction coefficients of P_r and Iⁱ_7(l() (= lumi-R) given by Eilfeld and Riidiger (1985, Z. Naturforsch. 40c , 109), significantly higher values are calculated for / as compared to φ found in the control experiments. These results can be explained either: (i) with a quantum yield ratio φ_pr-φ1700: φ_1700φpr=1:1 and an assumed additional dark reaction leading from Iⁱ₇₀₀ or later intermediates back to P_r: or (ii) with a quantum yield ratio φ_{pr φ 1700}: φ_{1700 φpr}=1:2. In this case all Iⁱ₇₀₀ have to relax to P_fr. In this case all Iⁱ₇₀₀ have to relax to P_fr.     

A PHYTOCHROME PHOTOTRANSFORMATION STUDY USING TWO-LASER/TWO-COLOR FLASH PHOTOLYSIS: ANALYSIS OF THE DECAY MECHANISM OF I700*

The mechanism of I₇₀₀ decay, representing an early event in the phytochrome P_r→ P_fr phototransformation, was reanalyzed in the microsecond range by conventional laser flash photolysis as well as by two-laser/two-color flash photolysis. Three kinetic models that might describe the I₇₀₀ decay mechanism following P_r excitation were considered: a parallel, a sequential, and an equilibrium model. These models were used to mathematically simulate both the one- and two-laser flash experiments in an effort to select the model best describing the I₇₀₀ decay. The sequential model could be excluded already on the basis of the one-laser flash photolysis results alone. Discussion of the two-laser/two-color flash rcsults in the context of the equilibrium and the parallel models is presented.     

THE EFFECT OF DEUTERIUM OXIDE ON THE FLUORESCENCE AND PHOTOTRANSFORMATION OF 124 kDALTON PHYTOCHROME*

Abstract— To probe the nature of primary photoprocess and the mechanism of the phototransformation of undegraded 124 kDa oat phytochrome, solvent deuterium isotope effects on the fluorescence and phototransformation of phytochrome have been investigated. The fluorescence intensity and lifetime of phytochrome (P_r form) are greater in D₂0-buffer than in H₂O-buffer, suggesting a possible involvement of proton transfer in the primary photoprocess of phytochrome. Although the photostationary equilibrium (P_r to P_fr ratio) was not altered by deuterium oxide, in contrast to degraded phytochrome, the rate constants of both transformations, P_r→ P_fr and P_fr→ P_r were enhanced by up to 24%. The P_r to P_fr phototransformation of degraded phytochrome, however, was retarded by about the same percentage in D₂O. These opposite effects of D₂O with degraded and undegraded phytochromes underscore the fact that the P_r form from the former reverts to the P_r form in the dark, apparently catalyzed by deuterated general and/or conjugate acidic group(s). With the degraded phytochrome the deuterium oxide enhancement of the rate of dark reversion was approximately 2-fold (Sarkar and Song, 1981). Both the fluorescence intensity and the rates of phototransformation of phytochrome were enhanced in D₂O with successive photocyclings (P_r→ P_fr→ P_r→ P_fr→ P_r etc.) with alternating red and far-red irradiation. It has been proposed that successive photocycling of phytochrome in D₂O results in proton-deuteron exchange in the partially exposed P_tr chromophore and/or its surrounding amino acid residues.     

Synthesis of nonlinear optical maleimide copolymer by polymer reaction and their electro-optic properties

Thermally stable poly(α-methyl styrene-co-maleimide) (MSMI) and poly(α-methyl styrene-co-4-carboxyphenyl maleimide) (MSCM) substrate polymers were obtained readily by free radical polymerization of comonomers. Introduction of a DR1 chromophore to the maleimide units of MSMI substrate polymer by the Mitsunobu reaction was dependent on the reaction solvent. The degree of substitution of DR1 into the MSMI polymer was bound to be 91.1 mol % and 0.4 mol % by UV spectrometers in the THF and DMF solvent, respectively. DR1 chromophore was, however, substituted in the MSCM polymer at 33.0 mol % by Mitsunobu reaction in the THF solvent. Both substrate and NLO polymer exhibited high thermal stability due to the incorporation of maleimide units in the polymer chain. The glass transition temperature (T_g) and initial decomposition temperature (T_i) of the NLO polymer were in the range of T_g = 185°C and T_i = 310–345°C. The electro-optic coefficient (r₃₃) of NLO polymer was determined with an experimental setup capable of the real-time measurement while varying both the poling field and temperature. The NLO polymer MSMI-THF had a higher r₃₃ value than MSCM-DR due to an increased degree of substitution of DR1 chromophore. MSMI-THF had a maximum r₃₃ value of 16 pm/V at 135 MV/m poling field with a 632.8 nm light source. © 1999 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 37: 3715–3722, 1999     

Mixed complex formation of lead(II) and mercury(II) ethylenediaminetetraacetates with thiourea in an aqueous solution

The formation of mixed-ligand complexes HgEdtaThio²⁻, HgEdtaS₂O₃⁴⁻, PbEdtaThio²⁻, and Pb(Thio) _i ²⁺, i = 1, 2; Thio is thiourea) was studied by calorimetry, pH metry, and ¹H and ¹³C NMR spectroscopy. The thermodynamic parameters (logK, Δ _r G ⁰, Δ _r H, and Δ _r S) for the formation of the complexes at 298.15 K and the ionic strength I = 0.5(NaClO₄) were determined. The most probable coordination mode of the ligands in the mixed complex was considered.     

Efficient Photoconversion Distorts the Fluorescence Lifetime of GFP in Confocal Microscopy: A Model Kinetic Study on Mutant Thr203Val

Phototransformations of autofluorescent proteins are applied in high‐resolution microscopy and in studying cellular transport, but they are detrimental when accidentally occurring in blinking or photobleaching (BL). Here, we investigate the kinetics of phototransformations of a photoactivatable green fluorescent protein (GFP) in confocal microscopy. Photoconversion (PC) is achieved by excitation of the barely present anionic chromophore state R_eq^? in the GFP mutant Thr203Val. Besides the shift of the equilibrium between the neutral chromophore state RH and R_eq^?, the photoconverted anionic chromophore R_PC^? exhibits a reduced fluorescence lifetime τ_fl=2.2 ns. In fluorescence lifetime imaging microscopy, τ_fl is found to depend, however, on the excitation conditions and history. The underlying photochemistry is described by the kinetic scheme of consecutive reactions, R_eq^?→R_PC^?→P_dark, in which the anionic chromophore species and the dark protein P_dark are coupled by PC and BL. Time‐correlated single‐photon‐counting detection in a confocal geometry of freely diffusing species is used to compute the quantum yields for PC and BL, Φ_PC and Φ_BL. The assessed values are Φ_PC=5.5×10^?4 and Φ_BL>1×10^?5. Based on these values, PC provokes misinterpretation in fluorescence resonance energy transfer experiments and is responsible for spectroscopic peculiarities in single‐molecule detection.     

A Cobalt(I) Pincer Complex with an η2‐Caryl−H Agostic Bond: Facile C−H Bond Cleavage through Deprotonation,Radical Abstraction,and Oxidative Addition

The synthesis and reactivity of a Co^I pincer complex [Co(ϰ³P,CH,P‐P(CH)P^NMe‐iPr)(CO)₂]⁺ featuring an η²‐ C_aryl−H agostic bond is described. This complex was obtained by protonation of the Co^I complex [Co(PCP^NMe‐iPr)(CO)₂]. The Co^III hydride complex [Co(PCP^NMe‐iPr)(CNtBu)₂(H)]⁺ was obtained upon protonation of [Co(PCP^NMe‐iPr)(CNtBu)₂]. Three ways to cleave the agostic C−H bond are presented. First, owing to the acidity of the agostic proton, treatment with pyridine results in facile deprotonation (C−H bond cleavage) and reformation of [Co(PCP^NMe‐iPr)(CO)₂]. Second, C−H bond cleavage is achieved upon exposure of [Co(ϰ³P,CH,P‐P(CH)P^NMe‐iPr)(CO)₂]⁺ to oxygen or TEMPO to yield the paramagnetic Co^II PCP complex [Co(PCP^NMe‐iPr)(CO)₂]⁺. Finally, replacement of one CO ligand in [Co(ϰ³P,CH,P‐P(CH)P^NMe‐iPr)(CO)₂]⁺ by CNtBu promotes the rapid oxidative addition of the agostic η²‐C_aryl−H bond to give two isomeric hydride complexes of the type [Co(PCP^NMe‐iPr)(CNtBu)(CO)(H)]⁺.     

A Cobalt(I) Pincer Complex with an η2‐Caryl−H Agostic Bond: Facile C−H Bond Cleavage through Deprotonation,Radical Abstraction,and Oxidative Addition

The synthesis and reactivity of a Co^I pincer complex [Co(?³P,CH,P‐P(CH)P^NMe‐iPr)(CO)₂]⁺ featuring an η²‐ C_aryl?H agostic bond is described. This complex was obtained by protonation of the Co^I complex [Co(PCP^NMe‐iPr)(CO)₂]. The Co^III hydride complex [Co(PCP^NMe‐iPr)(CNtBu)₂(H)]⁺ was obtained upon protonation of [Co(PCP^NMe‐iPr)(CNtBu)₂]. Three ways to cleave the agostic C?H bond are presented. First, owing to the acidity of the agostic proton, treatment with pyridine results in facile deprotonation (C?H bond cleavage) and reformation of [Co(PCP^NMe‐iPr)(CO)₂]. Second, C?H bond cleavage is achieved upon exposure of [Co(?³P,CH,P‐P(CH)P^NMe‐iPr)(CO)₂]⁺ to oxygen or TEMPO to yield the paramagnetic Co^II PCP complex [Co(PCP^NMe‐iPr)(CO)₂]⁺. Finally, replacement of one CO ligand in [Co(?³P,CH,P‐P(CH)P^NMe‐iPr)(CO)₂]⁺ by CNtBu promotes the rapid oxidative addition of the agostic η²‐C_aryl?H bond to give two isomeric hydride complexes of the type [Co(PCP^NMe‐iPr)(CNtBu)(CO)(H)]⁺.     

Reduction of iodine by hydroxylamine within the pH range 0.7–3.5

The reduction of iodine by hydroxylamine within the [H⁺] range 3×10⁻¹–3×10⁻⁴ mol.L⁻¹ was first studied until completion of the reaction. In most cases, the concentration of iodine decreased monotonically. However, within a narrow range of reagent concentrations ([NH₃OH⁺]₀/[I₂]₀ ratio below 15, [H⁺] around 0.1 mol.L⁻¹, and ionic strength around 0.1 mol.L⁻¹), the [I₂] and [I₃⁻] vs. time curves showed 2 and 3 extrema, respectively. This peculiar phenomenon is discussed using a 4 reaction scheme (I₂+I⁻⇔︁I₃⁻, 2 I₂+NH₃OH⁺+H₂O→HNO₂+4 I⁻+5 H⁺, NH₃OH⁺+HNO₂→N₂O+2 H₂O+H⁺, and 2 HNO₂+2 I⁻+2 H⁺→2 NO+I₂+2 H₂O). In a flow reactor, sustained oscillations in redox potential were recorded with an extremely long period (around 24 h). The kinetics of the reaction was then investigated in the starting conditions. The proposed rate equation points out a reinforcement of the inhibition by hydrogen ions when [H⁺] is above 4×10⁻² mol.L⁻¹ at 25°C. A mechanism based on ion-transfer reactions is postulated. It involves both NH₂OH and NH₃OH⁺ as the reducing reactive species. The additional rate suppression by H⁺ at low pH would be connected to the existence of H₂OI⁺ in the reactive medium. © 1998 John Wiley & Sons, Inc. Int J Chem Kinet 30: 785–797, 1998     

Simulation of the kinetics of adenosine 5′-triphosphate hydrolysis catalyzed by the Cu2+ ion: The role of conformation and the catalytic effect of the OH− ion

The hydrolysis kinetics of the dimeric complex (CuATP^2? · OH₂)₂ {D} up to ≈40% ATP conversion at 25°C, pH 5.7–7.8, and [Cu · ATP]₀ = (2.07 ± 0.03) × 10^?3 mol/l is analyzed by numerical simulation. CuADP^? + P_i (P_i is an inorganic phosphate) form from DOH^?, and the latter forms rapidly from D. The abstraction of H⁺ from the coordinated H₂O molecule is an irreversible reaction involving an OH^? ion from the medium. The maximum possible DOH^? concentration at a given pH is reached at the initial stage of hydrolysis (0.3–6.0 min after the initiation of hydrolysis). CuADP^? + P_i form from D via two consecutive irreversible steps. The ADP buildup rate in the process is determined by the reversible conformational transformation of DOH^? resulting in a pentacovalent intermediate (IntK). OH^? ions from the medium are involved both in IntK formation and in the reverse reaction and are a hydrolysis inhibitor. AMP forms from the intermediate IntK₃, which forms reversibly from DOH^?, OH^? ions from the medium being involved in the forward and reverse reactions. This is followed by irreversible (AMPH)^? formation involving H₃O⁺ ions from the medium. The rate and equilibrium constants are determined for the formation and decomposition of hydrolysis intermediates. The concentrations of the intermediates are plotted versus time for various pH values. The structures of the intermediates are suggested. The causes of a peak appearing in the initial ADP formation rate versus pH curve are analyzed.     

Quantum mechanics and quasiclassical study of the H/D+FO → OH/OD+F,HF/DF+O reactions: Chemical stereodynamics

The time‐dependent quantum wave packet and the quasi‐classical trajectory (QCT) calculations for the title reactions are carried out using three recent‐developed accurate potential energy surfaces of the 1¹A′, 1³A′, and 1³A″ states. The two commonly used polarization‐dependent differential cross sections, dσ₀₀/dω_t, dσ₂₀/dω_t, with ω_t being the polar coordinates of the product velocity ω′, and the three angular distributions, P(θ_r), P(Φ_r), and P(θ_r,Φ_r), with θ_r, Φ_r being the polar angles of the product angular momentum, are generated in the center‐of‐mass frame using the QCT method to gain insight into the alignment and the orientation of the product molecules. Influences of the potential energy surface, the collision energy, and the isotope mass on the stereodynamics are shown and discussed. Validity of the QCT calculation has been examined and proved in the comparison with the quantum wave packet calculation. © 2009 Wiley Periodicals, Inc. J Comput Chem, 2010     

Mass transfer from copper melts by top-blowing of an argon-hydrogen plasma jet

The volatilization of lead, copper, tin, and zinc from copper melts using the technique of top-blowing with an argon-hydrogen plasma jet was experimentally investigated and theoretically evaluated. A plasma burner with 16 kW power was used in the experiments. The mole flow of the plasma gases was 0.017 mol/s (25 liters/min when T = 25°C and P_G = 1 bar). The temperature was 1830°C on the surface of the melt and between 1200 and 1500°C in the molten solution. When the zinc concentration is above 2 mole%, supersaturation of zinc occurs on the surface. In this range of concentrations the ratio of dilution of the concentration in relation to time is linear (zeroth-order reaction). When the concentration of zinc is below 2 mole%, the time dependence of volatilization can be described by an exponential law corresponding to a first-order reaction, because in this case the rate-determining step is the mass transport of zinc in the molten copper phase. From the change from zeroth-order to first-order reaction during the volatilization of zinc, the temperature on the surface of the melt can be estimated with a high degree of accuracy. On the other hand, the volatilization of tin and lead is determined by mass transfer in the gas phase, which leads to an exponential law for the whole range of concentrations. Reaction models were set up on the basis of the experimental data. The relationships thereby obtained permit one to evaluate in advance the yield of future industrial volatilization processes with top-blown plasma jets.Nomenclature A activity - A _e , m² effective mass-transport or mass-transfer area - k, mol/m² s^–1 mass-transfer coefficient - k _g , mol/m² s^–1 mass-transfer coefficient in the gas phase - k _s , mol/m² s^–1 mass-transfer coefficient in the melt phase - k, mol/m² s^–1 overall mass-transfer coefficient - K =P _i /x _i equilibrium coefficient (distribution coefficient) - K _T =P _i /a _i equilibrium coefficient - n _s , mol number of moles of the melt phase - n _G , mol number of moles of the gas phase - n _g , mol/s mole flow of the top-blown gas - n _i , mol/s mole flow of the extract i into the gas phase - P _i , N/ M² partial pressure of extract substance i in the gas phase - t, s time - T, °C or K temperature: T_s, in the melt; T , at the phase interface - x _i , mol/mol concentration in the melt - X _f ¹ , mol/mol concentration in the melt at the phase interface - w, s^–1 rate constant - y _i , mol/mol concentration in the gas phase - y _f ⁱ , mol/mol concentration in the gas phase at interface - z, m coordinate in the direction of mass transfer - activity coefficient - ^O Henry coefficient     

Computational design of intermolecularly stabilized cyclic compounds with low-valent phosphorus

Understanding the structure, bonding, and stability of the P^I compounds has continued to be one of the very important research areas of low-valent main group chemistry. Up to now, many intramolecularly stabilized P^I in cyclic form and several intermolecularly stabilized P^I in acyclic form have been reported. In this paper, we report the computational design of two novel cyclic intermolecularly stabilized P^I types, i.e., RP(μ-R)AlR₂ 1 and P(μ-R)₂AlR₂ 2. For most of the 26 kinds of the studied substituents, both types of the P^I structures are more stable than the classical P^III-one R₂P-AlR₂ 3. The P^I feature is supported by the structural, natural bond orbital analysis, and reactivity analysis. Formally, 1 and 2 are cooperatively stabilized by two sets of intermolecular donor–acceptor interactions between AlR₃ and PR, i.e., μ-R → P and P → Al in 1 and μ-R₁ → P and μ-R₂ → Al in 2. These novel P^I compounds strongly welcome the future laboratory studies.     

STUDIES ON PHYTOCHROME PHOTOCONVERSIONS IN VITRO WITH LASER-INDUCED OPTOACOUSTIC SPECTROSCOPY*

After excitation at room temperature with a 15 ns dye laser pulse, phytochrome (60 kDalton) from etiolated oat seedlings was studied by optoacoustic spectroscopy, which records the heat emission caused by non-radiative deactivation processes of the photoexcited molecule. The action spectrum for the heat emitted by P_r deviates from the absorption spectrum around 610 and 695 nm, indicating the build-up of photoproduct(s) within 15 ns after excitation. It is proposed that the 695 nm product is identical with I₇₀₀ and lumi-R known to be the first intermediate on the P_r× P_fr pathway, and that the photochemical back reaction of I₇₀₀ to P_r occurs on the same time scale. About 90% of the absorbed light energy was lost by radiationless deactivation. Values for the rate constants of excited state deactivation of P_r and for the internal energy content difference of the primary photoproducts are calculated.     

Structural Insights into the Trp‐Cage Folding Intermediate Formation

The 20 residue long Trp‐cage is the smallest protein known, and thus has been the subject of several in vitro and in silico folding studies. Here, we report the multistate folding scenario of the miniprotein in atomic detail. We detected and characterized different intermediate states by temperature dependent NMR measurements of the ¹⁵N and ¹³C/¹⁵N labeled protein, both at neutral and acidic pH values. We developed a deconvolution technique to characterize the invisible—fully folded, unfolded and intermediate—fast exchanging states. Using nonlinear fitting methods we can obtain both the thermodynamic parameters (ΔH^F–I, T_m^F–I, ΔC_p^F–I and ΔH^I–U, T_m^I–U, ΔC_p^I–U) and the NMR chemical shifts of the conformers of the multistate unfolding process. During the unfolding of Trp‐cage distinct intermediates evolve: a fast‐exchanging intermediate is present under neutral conditions, whereas a slow‐exchanging intermediate‐pair emerges at acidic pH. The fast‐exchanging intermediate has a native‐like structure with a short α‐helix in the G¹¹–G¹⁵ segment, whereas the slow‐exchanging intermediate‐pair presents elevated dynamics, with no detectable native‐like residue contacts in which the G¹¹? P¹² peptide bond has either cis or trans conformation. Heteronuclear relaxation studies combined with MD simulations revealed the source of backbone mobility and the nature of structural rearrangements during these transitions. The ability to detect structural and dynamic information about folding intermediates in vitro provides an excellent opportunity to gain new insights into the energetic aspects of the energy landscape of protein folding. Our new experimental data offer exceptional testing ground for further computational simulations.     

二元无机物标准熵 So298的拓扑研究

为了预测二元无机物的标准熵,基于分子图的连接矩阵和离子参数gi、qi,提出了一种新的连接性指数mQ, mG及其逆指数mQ’, mG’。 qi、gi定义为：qi=(1.1+Zi1.1) /(1.7+ni), gi=(1.4+Zi) /(0.9+ri+ri-1),其中Zi 、ni和 ri分别代表离子i的电荷数、最外层主量子数和半径。从0Q, 0Q’, 1G,和1G’,利用多元线性回归分析方法和人工神经网络方法,可以构建优良的QSPR模型。对371个二元无机物,其多元线性模型及神经网络模型的相关系数、标准偏差和平均绝对偏差分别是：0.9905, 8.29 J.K-1.mol-1, 6.48 J. K-1.mol-1, 0.9960, 5.37 J.K-1.mol-1 和 3.90 J.K-1.mol-1。留一法交叉验证表明,其多元线性模型具有良好的稳定性。两个模型对187个未进入模型的二元无机物的标准熵的预测值和实验值之间的相关系数、标准偏差和平均绝对偏差分别是：0.9897, 8.64 J. K-1.mol-1, 6.84 J. K-1.mol-1, 0.9957, 5.63 J.K-1.mol-1, 和 4.18 J.K-1.mol-1。研究表明,本文方法在预测二元无机物标准熵时比文献方法更有效,两种模型均能较精确的预测二元无机物的标准熵,且神经网络模型的预测结果更精确。     

A Nickel Complex Containing a Pyramidalized,Ambiphilic Pincer Germylene Ligand

Reaction of N-heterocyclic carbene (NHC)-stabilized PGeP-type germylene Ge{o-(PiPr₂)C₆H₄}₂⋅^MeIiPr ( 1 ) (^MeIiPr=1,3-diisopropyl-4,5-dimethylimidazol-2-ylidene) with Ni(cod)₂ gave pincer germylene complex Ni[Ge{o-(PiPr₂)C₆H₄}₂](^MeIiPr) ( 2 ), in which the Ge center of 2 is significantly pyramidalized. Theoretical calculation on 2 predicted the ambiphilicity of the germanium center, which was confirmed by reactivity studies. Thus, complex 2 reacted with both Lewis base ^MeIMe (^MeIMe=1,3,4,5-tetramethylimidazol-2-ylidene) and Lewis acid BH₃⋅SMe₂ at the germanium center to afford the adducts Ni[Ge{o-(PiPr₂)C₆H₄}₂⋅^MeIMe](^MeIiPr) ( 3 ) and Ni[Ge{o-(PiPr₂)C₆H₄}₂⋅BH₃](^MeIiPr) ( 4 ), respectively. Furthermore, the former was slowly converted to dinuclear complex Ni₂[Ge{o-(PiPr₂)C₆H₄}₂]₂(^MeIMe)₂ ( 5 ) at room temperature. Complex 5 can be regarded as a dimer of the ^MeIMe analog of 2 with a Ni-Ge-Ge-Ni linkage.     

Synthesis,Structure, Reactivity and Catalytic Implications of a Cationic,Acetylide-Bridged Trigold–JohnPhos Species

The cationic complex [(JohnPhos–Au)₃(acetylide)][SbF₆] (JohnPhos=(2-biphenyl)di-tert-butylphosphine, L1) has been characterised structurally and features an acetylide–trigold(I)–JohnPhos system; the trinuclear–acetylide unit, coordinated to the monodentate bulk phosphines, adopts an unprecedented μ,η¹,η²,η¹ coordination mode with an additional interaction between distal phenyl rings and gold centres. Other cationic σ,π-[(gold(I)L1)₂] complexes have also been isolated. The reaction of trimethylsilylacetylene with various alcohols (iPrOH, nBuOH, n-HexOH) catalysed by cationic [Au^IL1][SbF₆] complexes in CH₂Cl₂ at 50 °C led to the formation of acetaldehyde acetals with a high degree of chemo- and regioselectivity. The reaction mechanism was studied, and several organic and inorganic intermediates have been characterised. A comparative study with the analogous cationic [Cu^IL1][PF₆] complex revealed different behaviour; the copper metal is lost from the coordination sphere leading to the formation of cationic vinylphosphonium and copper nanoparticles. Additionally, a new catalytic approach for the formation of this high-value cationic vinylphosphonium has been established.