Fluorescence quenching of anthracene by indole derivatives in phospholipid bilayers

The quenching of anthracene fluorescence by indole, 1,2-dimethylindole (DMI), tryptophan (Trp) and indole 3-acetic acid (IAA) in palmitoyloleoylphosphatidylcholine (POPC) lipid bilayers was investigated. A very efficient quenching of the anthracene fluorescence in the lipid membrane is observed. Stern-Volmer plots are linear for DMI but present a downward curvature for the other quenchers. This was interpreted as an indication of the presence of an inaccessible fraction of anthracene molecules. By a modified Stern-Volmer analysis the fraction accessible to the quenchers and the quenching constant were determined. The changes in the fluorescence emission spectrum of indole and DMI have been used to calculate the partition constants of these probes into the membranes, and bimolecular quenching rate constants were determined in terms of the local concentration of quencher in the lipid bilayer. The rate constants are lower than those in homogeneous solvents, which may be ascribed to a higher viscosity of the bilayer. No changes in the emission spectra of Trp and IAA are observed in the presence of vesicles, indicating that these probes locate preferentially in the aqueous phase, or in close proximity to the vesicular external interface in a medium resembling pure water. In these cases quenching rate constants were determined in terms of the analytical concentration. In the quenching by DMI a new, red shifted, emission band appears; it is similar to that observed in non-polar solvents and it is ascribable to an exciplex emission. The exciplex band is absent in the quenching by IAA and Trp and only very weakly present when the quencher is indole. From the position of the maximum of the exciplex emission, a relatively high local polarity could be estimated for the region of the bilayer where the quenching reaction takes place.     

Exciplex-type behavior and partition of 3-substituted indole derivatives in reverse micelles made with benzylhexadecyldimethylammonium chloride, water and benzene

The fluorescence properties of 3-methylindole (MI), 3-indoleacetic acid (IAA), 3-indoleethyltrimethylammonium bromide (IETA), L-tryptophan (Trp) and tryptamine hydrochloride (TA) were studied in reverse micelles solutions made with the cationic surfactant benzylhexadecyldimethylammonium chloride (BHDC) in benzene as a function of the molar ratio water/surfactant R (= [H2O]/[BHDC]). The fluorescence quenching of the model compound MI by benzene in cyclohexane solutions and by BHDC in benzene solutions were also studied in detail. The fluorescence of MI in benzene is characteristic of a charge-transfer exciplex. The exciplex is quenched by the presence of BHDC, due to the interactions of the surfactant ion pairs with the polar exciplex. In reverse micelle solutions at low R values, all the indoles show exciplex-type fluorescence. As R increases, the fluorescence behavior strongly depends on the nature of the indole derivative. The anionic IAA remains anchored to the cationic interface and its fluorescence is quenched upon water addition due to the increases of interface's micropolarity. For IETA, TA and Trp an initial fluorescence quenching is observed at increasing R, but a fluorescence recovery is observed at R > 5, indicating a probe partition between the micellar interface and the water pool. For the neutral MI, the fluorescence changes with R indicate the partition of the probe between the micellar interface and the bulk benzene pseudophase. A simple two-site model is proposed for the calculation of the partition constants K as a function of R. In all cases, the calculation showed that even at the highest R value, about 90% of the indole molecules remain associated at the micellar interface.     

Study of quenching of anthracene fluorescence by [60]fullerene

[60]Fullerene has been shown to have a very high quenching effect on the fluorescence of anthracene at room temperature in n-hexane, n-heptane and carbontetrachloride medium. The possibility that the quenching is due to ground state electron donor-acceptor (EDA) complex formation between [60]fullerene and anthracene has been shown to be untanable in the concentration range used ( approximately 10(-5)moldm(-3) in both anthracene and C(60)). No exciplex formation under the present experimental conditions has been observed. In the non-quenching solvents n-hexane and n-heptane the Stern-Volmer constant follows the right trend with respect to change in solvent viscosity but in case of the quenching solvent CCl(4), the trend is opposite.     

Emission from aromatic hydrocarbon-diene and -olefin exciplexes

Luminescence from aromatic hydrocarbon-olefin and -diene exciplexes, providing strong evidence for their intermediary in singlet quenching processes, is reported. Solvent dependence of the emission maximum gives a value of 10.8D for the dipole moment of the 1-cyanonaphthalene-1,2-dimethylcyclopentene exciplex while the temperature dependence affords a value of 6.7 kcal/M for the heat of formation. Linear-free-energy correlations of rate constants for 1-cyanonaphthalene and naphthalene fluorescence quenching by dienes and olefins and strained hydrocarbons with the adiabatic ionization potentials of the quenchers are consistent with major contributions from charge-resonance in the exciplex formation process.     

Studies on halogen quenching through the Stern-Volmer plot (author's transl)

The quenching effect for halogenated benzenes, methanes and ethanes have been investigated. The halogen quenching was accurately measured using the internal conversion electrons emitted from 113Sn-113mIn. From the quenching constants determined by the Stern-Volmer plots with respect to various halogen quenchers, the following results have been obtained. (1) The quenching constants increase with the number of halogen substituents, so as linearly in halogenated benzenes and exponentially in halogenated methanes and ehtanes. Even the isomers of halogenides have different quenching constants. (2) There is a linearity between logarithm of the quenching constant and a polarographic half-wave reduction potential. (3) Electron excitation provides larger quenching constants than UV excitation for halogenated methames. Based on these results, the mechanism of halogen quenching have been discussed in connection with the exciplex formation.     

Steady-state and time-resolved fluorescence studies on Trichosanthes cucumerina seed lectin

Steady-state and time-resolved fluorescence spectroscopic studies have been carried out on Trichosanthes cucumerina seed lectin (TCSL). The fluorescence emission maximum of TCSL in the native state as well as in the presence of 0.1 M lactose is centered around 331 nm, which shifts to 347 nm upon denaturation with 8 M urea, indicating that all the tryptophan residues of this protein in the native state are in a predominantly hydrophobic environment. The exposure and accessibility of the tryptophan residues of TCSL and the effect of ligand binding on them were probed by quenching studies employing two neutral quenchers (acrylamide and succinimide), an anionic quencher (I(-)) and a cationic quencher (Cs(+)). Quenching was highest with acrylamide and succinimide with the latter, which is bulkier, yielding slightly lower quenching values, whereas the extent of quenching obtained with the ionic quenchers, I(-) and Cs(+) was significantly lower. The presence of 0.1 M lactose led to a slight increase in the quenching with acrylamide and iodide, whereas quenching with succinimide and cesium ion was not significantly affected. When TCSL was denatured with 8 M urea, both acrylamide and succinimide yielded upward-curving Stern-Volmer plots, indicating that the quenching mechanism involves both dynamic and static components. Quenching data obtained with I(-) and Cs(+) on the urea-denatured protein suggest that charged residues could be present in close proximity to some of the Trp residues. The Stern-Volmer plots with Cs(+) yielded biphasic quenching profiles, indicating that the Trp residues in TCSL fall into at least two groups that differ considerably in their accessibility and/or environment. In time-resolved fluorescence experiments, the decay curves could be best fit to biexponential patterns, with lifetimes of 1.78 and 4.75 ns for the native protein and 2.15 and 5.14 ns in the presence of 0.1 M lactose.     

二烷基苯胺在饱和烷烃中对蒽和芘的荧光淬灭

研究了1,3-双(N-苯基-N-甲胺基)丙烷(DMA-AMD)、N-甲基-N-乙基苯胺(MEA)和二甲基苯胺(DMA)在正戊烷、甲基环己烷和正十六烷中对蒽和芘的荧光淬灭。荧光淬灭的同时有相应的激基复合物形成。发现DMA-AMD是一种比MEA或DMA都更为有效的淬灭剂,但生成的激基复合物的荧光效率很低。结果可用DMA-AMD内的另一个苯胺基与激基复合物产生分子内的淬灭来说明。这些苯胺的双分子淬灭速度常数取决于溶剂的粘度,而且在粘稠的烷烃中可以超过理论上扩散控制的限度。     

Tryptophan Modification and Fluorescence Spectrum of Hyaluronidase

Tryptophan residues in hyaluronidase （HAase） were modified by N-bromosuccinimide （NBS）, the results indicated that there were eleven tryptophan residues in HAase, one of which was exposed and essential for the activity of the enzyme. The study on fluorescence quenching showed that KI could not quench all of the fluorescence from Trp residues in HAase, while acrylamide （Acr） could quench almost all of the fluorescence from Trp residues in HAase. The collisional quenching constants （KD） of HAase at different concentrations of Acr were calculated in terms of Stern-Volmer equation. The results implied that some of the Trp residues were buried in the interior of HAase and the Trp residue on the surface of HAase was not located in the hydrophobic pocket.     

Porphinato Zinc Complexes Incorporated into the Bilayer Membrane of Lipid Liposomes

3,8,13,17-Tetramethyl-7,12-divinyl-2,18-bis(18-hydroxyoctadecyl propionate) porphinato zinc (BHPZn) and a model compound, dimethyl ester of protoporphinato zinc (DMPZn), were synthesized and incorporated into the hydrophobic region of bilayer membrane of dipalmitoylphosphatidylcholine (DPPC) liposome. The introduction of long alkyl groups onto the porphyrin ring is effective for restriction of porphyrin aggregation in the bilayer membrane of DPPC liposome. When the molar ratio of DPPC lipid to porphyrin is above 100, the spectrum of BHPZn in the liposome suggests that it is in a typically monomeric state. Quenching of BHPZn fluorescence in the hydrophobic bilayer membrane by hydrophilic quenchers is slow and shows smaller Stern-Volmer constants, while the quenching by hydrophobic quenchers shows much larger Stern-Volmer constants than that of the model compound, DMPZn. These results suggest that the location of the porphyrin ring of BHPZn is fixed at a certain depth in the hydrophobic bilayer membrane of DPPC liposome, and that that of DMPZn is widely distributed in the whole hydrophobic region.     

Photophysical properties of sexi(3-methoxythiophene): evidence for energy migration in a conducting oligomer, based on fluorescence quenching

The electronic absorption and fluorescence excitation and emission spectra of poly(3-methoxythiophene) (PMOT), a soluble, electroactive oligomer mainly constituted of hexamer, were investigated in dimethylsulfoxide at room temperature. The study of PMOT photophysical properties suggests the existence of important pi-electronic delocalization in the oxidized and reduced form of the oligomer. A significant fluorescence quenching of PMOT takes place in the presence of various quenchers such as dimethylterephthalate, potassium iodide and thallium acetate. Quenching mechanisms implying a quencher-induced intersystem-crossing step are proposed. Modified Stern-Volmer relationships were obtained with large quenching bimolecular rate constants (2.7 x 10(9)-6.1 x 10(11) l mol(-1) s(-1)), which suggests electronic energy migration throughout the repeat units of the PMOT hexamer.     

Interaction between a novel intramolecular charge transfer fluorescent probe PFEP and human serum albumin

<正>The interaction mechanism between human serum albumin(HSA) and 1-phenyl-3(fluorenone-2-yl)-5-(9-ethylcarbazole-3-yl)-2- pyrazoline(PFEP) was investigated by fluorescence and absorption titration techniques in combination with molecular modeling method.Stern-Volmer plots at different temperatures proved that PFEP could quench the intrinsic fluorescence of HSA attributed to a static quenching procedure.The association constants were calculated in the range of 1×10~5-8×10~5mol~(-1) at different pH conditions,and the stoichiometric ratio of binding was 1:1 between PEEP and HSA.Molecular modeling study showed that the distance between indole moiety of the Trp214 residue and the carbazole group at the terminal of PFEP was 4.45 A in the optimal model.     

ACRYLAMIDE QUENCHING OF TRYPTOPHAN PHOTOCHEMISTRY AND PHOTOPHYSICS

Studies of acrylamide quenching of tryptophan (Trp) fluorescence, photochemistry, and photoionization have been conducted. Quenching of Trp fluorescence in aqueous solution by addition of acrylamide in the concentration range 0.0-0.5 M was measured and resulted in a Stern-Volmer quenching constant of KSV = 21 +/- 3 M-1. Photolysis experiments were performed in which Trp was photolyzed at 295 nm in the presence of varying concentrations of acrylamide. The loss of Trp was monitored using reverse-phase high performance liquid chromatography (RP-HPLC) and was observed to follow first order kinetics. Production of N-formylkynurenine (NFK) was observed by RP-HPLC in irradiated Trp samples both in the presence and absence of added acrylamide. In addition, no new photochemical product was detected. This was taken as evidence that acrylamide did not alter the photochemical pathway but just reduced the reaction rate as expected for a physical quenching mechanism. Plotting the reciprocal of photolysis rate constant versus acrylamide concentration produced a Stern-Volmer constant for quenching of Trp photochemistry of KSV = 6 +/- 2 M-1. The KSV values for both fluorescence quenching and photolysis quenching were thus large, implying efficient quenching of both processes by acrylamide. Assuming an excited singlet state lifetime of 2.8 ns, the calculated second-order quenching rate constants for fluorescence and photolysis were kq = 7.5 x 10(9) and 2.1 x 10(9) M-1 s-1 respectively. The possible involvement of photoionization in the photolysis mechanism was investigated by studies of acrylamide quenching of voltage transients produced by xenon flash lamp excitation of Trp at aqueous/teflon or aqueous/mica interfaces.(ABSTRACT TRUNCATED AT 250 WORDS)     

在不同激发波长下的单壁碳纳米管的拉曼光谱研究

采用直流电弧法制备单壁碳纳米管样品,用457.5和632.8nm两种不同的激发光分别测得单壁碳纳米管的正常拉曼光谱和共振拉曼光谱.通过理论分析得到了单壁碳纳米管的直径分布,进一步推测了其类型及结构参数;对单壁碳纳米管的正切拉伸模的成分进行了归属.在632.8nm激发波长下得到了IG/ID值随激光功率变化的曲线,认为在2.5mW时,单壁碳纳米管缺陷的结构可能发生了改变.在用457.5nm波长激发的单壁碳纳米管的拉曼光谱中,首次发现了1421cm-1的拉曼谱峰.     

Photophysical processes of a copolymer containing naphthalene and carbazole rings

The photophysical processes of copolymer formed by copolymerization of beta-naphthyl-methacrylate (NMA) with vinylcarbazole (VCZ) were studied. The results show that when the solution of copolymer (NMA-VCZ) in THF is located in a low concentration range (about 10(-8) mol/l), the fluorescence emission is in good agreement with that of NMA monomer and the excimer is formed with gradual increase in concentration of copolymer (NMA-VCZ). The fluorescence of copolymer (NMA-VCZ) can be efficiently quenched both by electron donors and acceptors where the quenching effects follow the Stern-Volmer equation. The dimolecular exciplex between copolymer (NMA-VCZ) and N,N-dimethylaniline (DMA) is formed and the triple exciplex is also observed in the same system.     

菊粉酶中色氨酸残基的化学修饰及其荧光光谱

用N-溴代琥珀酰亚胺为修饰剂, 研究菊粉酶中的Trp残基的分布及其对酶生物学功能的影响, 为进一步研究菊粉酶结构与其功能之间的关系提供了新的信息.     

FLUORESCENCE QUENCHING OF INDOLIC COMPOUNDS IN REVERSE MICELLES OF AOT

Abstract— The fluorescence properties of indole, 1,2-dimethylindole and tryptophan have been determined in sodium bis(2-ethylhexyl)sulfosuccinate inverse micelles in heptane. Fluorescence quenching studies were done using CCl₄ and acrylamide as quenchers localized in the organic and micellar pseudophase respectively. The Stern-Volmer plots show departure from linearity and are dependent on the emission wavelength. The results were interpreted using a three pseudophases model. 1,2-Dimethylindole is predominantly solubilized in the n -heptane; however a small interaction with the micelles is apparent at high detergent concentration. Indole is distributed between the organic and the interfacial microphase with a partition constant of 34 ± 3, independent of the water content. Tryptophan is entirely localized at the interface. However the quenching experiments show that tryptophan molecules are not uniformly distributed in the interfacial pseudophase.     

Determination of environmentally important metal ions by fluorescence quenching in anionic micellar solution

This work describes the effect of a variety of metal ions as quenchers of the fluorescence of naphthalene, in aqueous micellar solutions of sodium dodecyl sulfate (SDS). The quenching by the metal ions can be adequately described by the Stern-Volmer equation and the best signal to noise ratios are obtained with low micellized detergent concentrations. Apparent Stern-Volmer constants decrease in the order: Fe3+ > Cu2+ > Pb2+ > Cr3+ > Ni2+ and directly reflect the relative sensitivity of the method for these ions. Detection limits (defined as three times the standard deviation of the blank for n= 10) for the fluorescence quenching of naphthalene by the metal ions in aqueous micellar SDS are in the range of 1.0 x 10(-6) to 1.0 x 10(-5) mol dm(-3). The proposed fluorescence quenching method shows good repeatibility for a variety of added quencher metal ions, indicating that anionic micelle-enhanced fluorescence quenching by metal ions constitutes an analytical method of rather general application.     

N-甲基-N-(2-甲基丙烯酰氧乙基)苯胺与其聚合物的荧光光谱

<正> 我们曾报道过甲基丙烯酸-4-N,N-二甲氨基苄酯(DMABMA)、N-(4-N′N′-二甲氨基苯基)代丙烯酰胺(DMAPMA)、8-丙烯酰氧喹啉(AQ)、N-丙烯酰-N′-苯基哌嗪(APP)等在同一分子中含有缺电子双键和给电子发色基团的单体及它们的聚合物在溶液中的荧光行为。在相同的链节克分子浓度下,这些单体的荧光强度比其聚合物的荧光强度低很多。我们将这种现象称为“结构自猝灭现象”。这种现象是由于共存在这类     

透明质酸酶的色氨酸残基修饰与荧光光谱研究

北五味子[Schisandra chinensis（Turcz．）Baill．]属广义木兰科植物,主产于我国东北,故又称“辽五味”,中药五味子的主要药材为北五味子的干燥果实,作为一种传统中药,五味子具有收敛固涩,益气生津,补肾宁心的功效,用于肺喘虚咳,心悸失眠诸病。     

ROLE OF THE TRYPTOPHAN FLUORESCENT STATE IN THE ULTRAVIOLET-INDUCED INACTIVATION OF β-TRYPSIN*

Abstract— Stern-Volmer quenching constants for β-trypsin at pH 3 were determined for fluorescence quenching by histidine, acrylamide, and nitrate ion. A modified Stern-Volmer plot (Lehrer, 1971) was employed to show that all of the fluorescent tryptophanyl residues of β-trypsin were equally susceptible to quenching by acrylamide at pH 3 when the enzyme was either in its native conformation or denatured in 6 M guanidine hydrochloride (GuHCl). Fluorescence lifetime measurements indicated that acrylamide quenched β-trypsin fluorescence by a purely collisional mechanism. Solvation of tryptophanyl residues of the protein was maximal at 2.5 M GuHCl, as monitored by fluorescence emission wavelength.
Investigations of the ultraviolet-induced inactivation of β-trypsin at 295 nm were performed in the presence of acrylamide at pH 3. The quantum yields for enzyme inactivation and indole destruction (determined using the PDAB reagent) were unchanged upon depopulation of the fluorescent state by 65 per cent, whether the enzyme was in its native conformation or denatured by 6 M GuHCl. It is concluded that the fluorescent state of tryptophanyl residues of β-trypsin is not involved in enzyme inactivation or tryptophan destruction.