Photocycle of 13-cis-retinal in bacteriorhodopsin caused by destruction to the cooperative effect of purple membrane

Ｉｎｔｅｇｒａｌｍｅｍｂｒａｎｅｐｒｏｔｅｉｎｓａｒｅｃｒｉｔｉｃａｌｔｏａｗｉｄｅｖａｒｉｅｔｙｏｆｃｅｌｌｕｌａｒｐｓｙｃｈｏｌｏｇｉｃａｌａｃｔｉｖｉｔｉｅｓ,ｓｕｃｈａｓｃｙｔｏｃｈｒｏｍｅｏｘｉｄａｓｅｉｎｍｅｄｉａｔｉｎｇｅｌｅｃｔｒｏｎｔｒａｎｓｐｏｒｔａｎｄｔｈｅａｃｔｉｖａｔｉｏｎｏｆｉｎｔｒａｃｅｌｌｕｌａｒｓｉｇｎａｌｓｂｙｔｒａｎｓｍｅｍｂｒａｎｅｒｅｃｅｐｔｏｒｓｃｏｕｐｌｅｄｔｏＧｐｒｏｔｅｉｎｓ[1].Ｉｎｔｈｅｓｅｍｅｍｂｒａｎｅａｃｔｉｖｉｔｉｅｓ,ｔｈｅｍｅｃｈａｎ…     

Optical properties of Triton X-100-treated purple membranes embedded in gelatin films

The purple membrane (PM) of the microorganism Halobacterium salinarium contains a hexagonally packed monolayer of the light-sensitive protein, bacteriorhodopsin (BR). The optical characteristics of gelatin-immobilized PMs depend strongly on the chemical environment of the PMs in the matrix. Here we present photoinduced absorptive and holographic characteristics of gelatin-embedded PMs solubilized with the non-ionic detergent, Triton X-100. The BR/detergent interaction was shown to slow the M-to-initial state transition of the photocycle and to increase the photosensitivity of the BR films. The lifetime of the holographic grating in Triton X-100-treated BR films was 2–3 times greater, when compared to the unmodified sample. Holographic grating growth times in BR films were shown to change depending on the extent of solubilization. The measured holographic sensitivity appeared to maximize in the range of Triton X-100/BR molar ratios from 15:1 to 25:1. The possible advantages of solubilized PM films as they are applied to optoelectronic devices are discussed.     

The laser-induced blue state of bacteriorhodopsin: mechanistic and color regulatory roles of protein-protein interactions, protein-lipid interactions, and metal ions

In this paper we characterize the mechanistic roles of the crystalline purple membrane (PM) lattice, the earliest bacteriorhodopsin (BR) photocycle intermediates, and divalent cations in the conversion of PM to laser-induced blue membrane (LIBM; lambda(max)= 605 nm) upon irradiation with intense 532 nm pulses by contrasting the photoconversion of PM with that of monomeric BR solubilized in reduced Triton X-100 detergent. Monomeric BR forms a previously unreported colorless monomer photoproduct which lacks a chromophore band in the visible region but manifests a new band centered near 360 nm similar to the 360 nm band in LIBM. The 360 nm band in both LIBM and colorless monomer originates from a Schiff base-reduced retinyl chromophore which remains covalently linked to bacterioopsin. Both the PM-->LIBM and monomer-->colorless monomer photoconversions are mediated by similar biphotonic mechanisms, indicating that the photochemistry is localized within single BR monomers and is not influenced by BR-BR interactions. The excessively large two-photon absorptivities (> or =10(6) cm(4) s molecule(-1) photon(-1)) of these photoconversions, the temporal and spectral characteristics of pulses which generate LIBM in high yield, and an action spectrum for the PM-->LIBM photoconversion all indicate that the PM-->LIBM and Mon-->CMon photoconversions are both mediated by a sequential biphotonic mechanism in which is the intermediate which absorbs the second photon. The purple-->blue color change results from subsequent conformational perturbations of the PM lattice which induce the removal of Ca(2+) and Mg(2+) ions from the PM surface.     

The effect of lipid environment in purple membrane on bacteriorhodopsin

The decay rate of the Bacteriorhodopsin (BR) photocycle intermediate M412 and proton, the proton pump efficiency (H+/M412), the ratios of M412 to other intermediates and the rotational correlation time (tauc) in purple membrane (PM) fragments treated by the zwitterionic detergent 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) with different concentrations were studied. The results show that: (1) The largest effect of CHAPS on M412 decay rate and proton decay rate of BR, tauc of PM and the ratios of M412 to other intermediates in BR photocycle is in the range of its critical micelle concentration (CMC). This indicates that changes of the ratios of M412 to other intermediates, tauc, M412 decay and proton decay occur and are due to the variation of the lipid environment. (2) The dependency of proton yield on CHAPS concentrations is basically consistent with that of M412s%. This indicates the relation between proton pumping function and M412. These studies show the importance of maintaining a native environment.     

The effect of some organic surfactants upon the reduction rate of In3+ in perchloric medium on mercury

The effect of Triton X-100, dodecylammonium, tetrabutylammonium, and dodecylsulphate upon the reduction rate of In³⁺ to indium amalgam in 0.1 M HClO₄ was investigated, both in the presence and in the absence of 1 M NaClO₄. After having accounted for the blocking effect upon the kinetics of In³⁺ reduction via the factor (1??), where ? is the surface coverage by the surfactant, the remaining electrostatic effect was compared with that predicted by the Frumkin theory for diffuse-layer effects. For this purpose the Gouy-Chapman-Stern theory was used to calculate the potential _d at the outer Helmholtz plane from measured values of the charge density q_M on the metal and of the charge density q_i due to the adsorbed surfactant. With the exception of Triton X-100, the electrostatic effect predicted by the Frumkin theory combined with the Gouy-Chapman-Stern theory was found to be much greater than that observed experimentally. This result was explained by the inadequacy of the Gouy-Chapman-Stern model of the double layer in the presence of bulky surface-active ions.     

Cloud point extraction of iron(III) and vanadium(V) using 8-quinolinol derivatives and Triton X-100 and determination of 10(-7)moldm(-3) level iron(III) in riverine water reference by a graphite furnace atomic absorption spectroscopy

The cloud point extraction behavior of iron(III) and vanadium(V) using 8-quinolinol derivatives (HA) such as 8-quinolinol (HQ), 2-methyl-8-quinolinol (HMQ), 5-butyloxymethyl-8-quinolinol (HO₄Q), 5-hexyloxymethyl-8-quinolinol (HO₆Q), and 2-methyl-5-octyloxymethyl-8-quinolinol (HMO₈Q) and Triton X-100 solution was investigated. Iron(III) was extracted with HA and 4% (v/v) Triton X-100 in the pH range of 1.70-5.44. Above pH 4.0, more than 95% of iron(III) was extracted with HQ, HMQ, and HMO₈Q. Vanadium(V) was also extracted with HA and 4% (v/v) Triton X-100 in the pH range of 2.07-5.00, and the extractability increased in the following order of HMQ < HQ < HO₄Q < HO₆Q. The cloud point extraction was applied to the determination of iron(III) in the riverine water reference by a graphite furnace atomic absorption spectroscopy. When 1.25 × 10⁻³ M HMQ and 1% (v/v) Triton X-100 were used, the found values showed a good agreement with the certified ones within the 2% of the R.S.D. Moreover, the effect of an alkyl group on the solubility of 5-alkyloxymethyl-8-quinolinol and 2-methyl-5-alkyloxymethyl-8-quinolinol in 4% (v/v) Triton X-100 at 25 °C was also investigated.     

A Photochemical Study of Uranyl Ion Interaction with the Triton X-100 Micellar System

This is a report on the spectroscopic characteristics of UO²⁺₂in the excited state in Triton X-100 micellar medium. It also indicates some important results of viscosity and surface tension measurements of the system which have direct relevance to the spectroscopic investigation in the excited state. The quenching of the UO²⁺₂fluorescence due to Triton X-100, upon micellization in the aqueous medium, reveals two kinds of microenvironments of the fluorophore from the Stern–Volmer plot. This has been verified by flash photolytic measurements. A blue shift of the quenched emission spectrum is ascribed to the collisional encounter of UO²⁺₂with the head groups of Triton X-100.     

INTERACTIONS OF BOTH MELITTIN AND ITS SITE-SPECIFIC MUTANTS WITH BACTERIORHODOPSIN OF Halobacterium halobium: SITES OF ELECI'ROSTATIC INTERACI'ION ON MELITI'IN

Abstract Melittin and its site-specific mutants differentially delay the slow-decaying component of the photocycle intermediate M₄₁₂ of bacteriorhodopsin in the purple membrane and the acetylated purple membrane whose several lysine residues are modified. This effect is attributed to the interaction of the total positive charges of melittin or its mutants with the total negative charges of bacteriorhodopsin. The effects of melittin and its mutants on the Triton X-100–solubilized bacteriorhodopsin monomers are somewhat complicated but are associated with their charges. These results show that there is electrostatic interaction between bacteriorhodopsin and melittin and that both N-and C-termini of melittin function as sites of the interaction, with Arg 22 and Arg 24 making a prominent contribution to the effective surface charge of melittin. Melittin, at certain concentrations, partially restores the decreased photoactivity of the bacteriorhodopsin monomers trapped in the Triton-lipid-protein mixed micelles, which suggests that melittin may compete with Triton X-100 for the binding sites on the bacteriorhodopsin monomers. Other kinds of interactions between bacteriorhodopsin and melittin are also indicated. The possible states of melittin in membranes are discussed.     

The Photostabilization of Hemoglobin in Triton X-100 Aqueous Solution and Triton X-100/n-C5H11OH/H2O Microemulsion

The influence of UV-irradiation on the interaction of hemoglobin (Hb) with Triton X-100 is investigated by UV–Vis absorption spectroscopy, fluorescence spectroscopy and freeze-fractured transmission electron microscopy. It is found that in Triton X-100/H₂O systems Hb can convert to hemichrome but heme is not present, whereas in Triton X-100/n-C₅H₁₁OH/H₂O microemulsion Hb can convert to hemichrome and then induce the heme monomer to leave the hydrophobic cavity of Hb. UV-irradiation can also convert Hb to hemichrome, and subsequently make heme to be photodegraded, in which the conversion rate depends on the structure of the surfactant aggregates. Furthermore, in order to understand the mechanism of photostabilization of Hb in Triton X-100 systems, the photostabilization of heme in the Triton X-100 aqueous solutions and Triton X-100/n-C₅H₁₁OH/H₂O microemulsions has been studied.     

Wetting and adsorption: I. The structure of adsorbed layer of nonionic surfactants at SiO2/water and SiO2/cyclohexane interface

Based on the adsorption of Triton X-100 on silica/water and silica/cyclohexane interfaces and the adsorption of Triton X-305 on silica/water interface, two adsorption models have been proposed. On silica/cyclohexane interface, the adsorption of Triton X-100 is monomolecular layer. The molecules in the monolayer are presumed to be attached to the silica surface by their EO chain such that their hydrocarbon chain are exposed to the cyclohexane phase. On silica/water interface, the adsorption of Triton X-100 or Triton X-305 is bimolecular layer. The surfactant molecules orientated in the first layer are similar with that on the silica/cyclohexane interface. The molecules in the second layer are postulated to adsorb on those of the first in the opposite orientation, with EO chain directed toward the adsorption medium. The contact angle of quartz-water-cyclohexane (θ_W) as a function of the concentration of Triton X-100 and Triton X-305 in water has been measured with quartz plate employing the captive drop (cyclohexane) technique. The observed θ_W (measured through water) rose from < 10° to a maximum of about 120° for Triton X-100 and of about 40° for Triton X-305 as the concentration of surfactant in water increased, and then fell, as the concentration increased further. The results are consistent with the proposed adsorption models.     

Study on cloud points of Triton X-100-cationic gemini surfactants mixtures: A spectroscopic approach

This study investigates the effects of various cationic surfactants on the cloud point (CP) of the nonionic surfactant Triton X-100 (TX-100) in aqueous solutions. Instead of visual observation, a spectrophotometer was used for measurement of the cloud point temperatures. The values of CPs for Triton X-100 can be measured directly because TX-100 has an average number of oxyethylene units per molecule of p ≈ 9.5 and a CP = 66.0 °C. Quaternary ammonium dimeric surfactants (m-s-m, m = 10, 12, and 16, and s = 2, 6, and 10) were synthesized and used. The melting temperature T_M and the Krafft temperature T_K were measured for 1 wt% aqueous solutions of these synthesized surfactants. The melting temperature of the solid gemini surfactants increased with the carbon number of the alkyl chain. The results showed that additions of the gemini surfactants (which are infinitely miscible with water) to Triton X-100 increased the cloud point of the TX-100 solutions. All salts tested in these studies had a large effect on the CPs of nonionic surfactants due to their effect on water structure and their hydrophilicity. The effect of the alkyl chain length of the gemini surfactant on the CP of Triton X-100 is therefore more important than the spacer chain length.     

Influence of the preparation conditions on the textural properties andn-hexane isomerization activity of Pt/SO42??ZrO2

The effect of Na₂SO₄ and PhCOONa on the aquation of [Fe(Me₄phen)₃]²⁺ has been investigated in pure Triton X-100 as solvent. The rate and mechanism of the aquation are explained in terms of changes in the mobility, activity and structure of H₂O in the restricted environment of water pockets in the Triton X-100 solvent.     

Modification of human erythrocyte membrane potentials by the action of the polyoxyethylene detergent Triton X-100

Triton X-100 in sublytic concentrations causes a marked increase in human erythrocyte membrane permeability to cations with small selectivity to K⁺ ions. This membrane permeabilization induces modification of membrane potential: hyperpolarization in low K⁺ media and depolarization in high K⁺ media. Since the specificity of the K⁺-ionophorous action of Triton X-100 is very low in comparison with valinomycin, the membrane potential changes are only in the range of −9 to +5 mV. It is suggested that they are determined by the generation of a diffusion potential across the membrane, although the influence of Triton X-100 on the potentials at cell surfaces cannot be neglected.     

A Study on the Location and Aggregation of Tetrahydrophenylporphyrin with a Single Hexadecyl Chain in Triton X-100 Micelle

The location and aggregation of 5,10,15-tris(4-hydroxyphenyl)-20-(hexadecyloxyphenyl)porphyrin (P) in nonionic polyoxyethylene (9.5) octylphenol (Triton X-100) micelle solutions were studied by means of UV–Vis and fluorescence spectra. P forms premicelle surfactant–porphyrin aggregates when the surfactant concentration is below and approaching the CMC. In Triton X-100 micelle solutions, different types of H-aggregates of P were formed when the concentration of P is higher than 3.9×10^-6?mol?dm^-3. As the bulk pH is changed, a transfer process for the porphyrin moiety in Triton X-100 micelle occurs. In neutral Triton X-100 micelle solutions, P may be located at the inner layer of the micelle; in basic conditions, the porphyrin moiety may transfer to the outer surface of the micelle. The kinetic study of porphyrin complexed with Cu(II) in Triton X-100 micelle solutions shows that the metalation rate could be controlled by changing the pH.     

All-trans to 13-cis retinal isomerization in light-adapted bacteriorhodopsin at acidic pH

The flash photolysis kinetic spectra of the intermediate M(412) of bacteriorhodopsin were monitored during the process of acid titration. In the light-adapted state, the maximum peak amplitude of M(412) absorbance of bacteriorhodopsin decreased (pK(a)=3.40+/-0.05) as the pH decreased from 7.3 to 1.9. In the dark-adapted state, the maximum peak amplitude of M(412) absorbance of bacteriorhodopsin increased as the pH decreased from 6.9 to 4.1, and then decreased (pK(a)=2.85+/-0.05) as the pH dropped to 2.1. These different trends in the change in the maximum peak amplitude suggested that not only the transition of purple membrane to blue membrane had taken place in both light and dark-adapted states, but also the fraction of all-trans-bR had changed during the acid titration. The pH-dependent absorption changes at 640 nm of bacteriorhodopsin in both light- and dark-adapted states were also observed. The pK(a)-values of the purple-to-blue transition were 3.80+/-0.05 in light-adapted state and 3.40+/-0.05 in dark-adapted state, respectively. According to Balashov's method, the fraction of all-trans-bR was assayed as the pH decreased. All these results indicated that the purple-to-blue transition of light-adapted bacteriorhodopsin was accompanied by an all-trans to 13-cis retinal isomerization at acidic pH.     

Collection of iron(III) from homogeneous aqueous solutions on membrane filters using Chromazurol B with Triton X-100.

A membrane filtration method was examined concerning the effective collection of iron(III) from a homogeneous aqueous solution with Chromazurol B (CAB), one of the triphenylmethane dyes, as a precipitating reagent in the presence of a non-ionic surfactant, polyethylene glycol mono[4-(1,1,3,3-tetramethylbutyl)phenyl]ether (Triton X-100). A formed blue Fe(III)-CAB complex was collected as a precipitate on a membrane filter by filtration under suction from a homogeneous aqueous solution in the pH range over about 2. The original solution was prepared at a concentration ratio of CAB to Fe(III) of to 10, and that of Triton X-100 to CAB of 10 to 100. It was then adjusted to a pH value of between 1.0 and 6.5. A linear relationship (r = 0.999) was obtained between the initial concentration and the found one of Fe(III) in the range of 2.0 x 10(-5) to 4.0 x 10(-4) mol dm(-3) at a fixed concentration ratio of CAB to Fe(III) of 3 and that of Triton X-100 to CAB of 20. This membrane filtration with CAB and Triton X-100 may be utilized for the separation of Fe(III) as a background species.     

Tensammetry with accumulation on the hanging mercury drop electrode : Part 3. The behaviour of triton X-100 in mixtures with PEG-9000

The behaviour of Triton X-100, which can be present in monomeric or associated form, and its mixtures with PEG-9000, which does not undergo association, is described. The tensammetric curve of Triton X-100 alone shows one or two peaks at negative potentials, depending on the concentration of Triton X-100, i.e., on the presence of associated forms. For <2 mg l^?1, there is one broad peak, related to monomers of Triton X-100. The calibration plot for this peak is sigmoidal but its rising section (0.05–0.20 mg l^?1) is approximately linear. The calibration curve of the second, much narrower, peak related to associated forms of Triton X-100, grows parabolically with increasing concentration of Triton X-100. The behaviour of a mixture of PEG-9000 with a larger amount of Triton X-100 is similar to the behaviour of a model mixture of components with sufficiently different properties (e.g., PEG 1500/PEG 9000). The peak for PEG-9000, the stronger surfactant, is relatively less affected by a large amount of Triton X-100. Even this effect can be decreased by using a suitable preconcentration potential (?1.45 V vs. SCE) so that PEG-9000 can be determined in the presence of a 1000-fold amount of Triton X-100. Both peaks of Triton X-100 are greatly decreased by the presence of PEG-9000 and the broad peak can be completely suppressed. Triton X-100 cannot be determined accurately in the presence of unknown amounts of PEG-9000.     

Determination of trace alpha-fetoprotein variant by affinity adsorption solid substrate-room temperature phosphorimetry and its application to the forecast of human diseases

In the presence of ion perturber LiAc, 4-generation polyamidoamine dendrimers (4G-D) could emit strong and stable room temperature phosphorescence (RTP) signal at on nitrocellulose membrane (NCM), and Triton X-100 could sharply enhance the RTP signal of 4G-D. Triton X-100-4G-D was used to label concanavalin agglutinin (Con A) to get the labeling product Triton X-100-4G-D-Con A. Quantitative specific affinity adsorption (AA) reaction between Triton X-100-4G-D-Con A and α-fetoprotein variant (AFP-V) could be carried out on the surface of NCM, whose product Triton X-100-4G-D-Con A-AFP-V could emit strong and stable RTP and its ΔI_p was proportional to the content of AFP-V. According to the facts above, a new affinity adsorption solid substrate-room temperature phosphorimetry (AA-SS-RTP) for the determination of trace AFP-V by Con A labeled with Triton X-100-4G-D was established. Detection limits of this method were 0.23 fg spot⁻¹ (direct method, corresponding concentration: 5.8 × 10⁻¹³ g mL⁻¹) and 0.13 fg spot⁻¹ (sandwich method, corresponding concentration: 3.2 × 10⁻¹³ g mL⁻¹). It has been successfully applied to determine the content of AFP-V in human serum and forecast human diseases, for its high sensitivity, long RTP lifetime, good repeatability, high accuracy and little background perturbation with at the long wavelength area. Meanwhile, the mechanism for the determination of trace AFP-V using AA-SS-RTP was also discussed.     

Surface Property of Nonionic Surfactant Triton X-100 in an Ionic Liquid

A common nonionic surfactant Triton X-100 was dissolved in a commercial ionic liquid (IL) 1-butyl-3-methylimidazolium tetrafluoroborate (bmimBF₄). The surface tension of the bmimBF₄ solution was decreased with increasing the content of surfactant Triton X-100, a similar phenomenon with aqueous solution systems. Dynamic surface properties of Triton X-100 in bmimBF₄ were measured. It was found that pure IL solvents need rearrangement at the air-bmimBF₄ interface during the beginning stage of absorption. Moreover, the adsorption model was found to be in accord with the diffusion-controlled adsorption mechanism, and further, the dilute bmimBF₄ solutions are close to the diffusion-controlled adsorption.     

Effects of Triton X-100 on Properties of Hemoglobin and Controlled Release of Drug in Hemoglobin/Ribavirin/H2O System

Effects of Triton X-100 on the properties of hemoglobin (Hb) and on the controlled release of ribavirin were studied using the methods of UV-Vis spectrum, fluorescence spectrum, zeta potential, conductivity, high-performance liquid chromatographic (HPLC), and image morphology in Hb/ribavirin/H₂O system. With the increase of concentration of Triton X-100 in the system, the intrinsic fluorescence intensity, synchronous fluorescence intensity, fluorescence polarization, zeta potential, and morphology of Hb all changed gradually, and the ribavirin located on the Hb surface was dissociated and released out. When the concentration of Triton X-100 was higher than 1×10⁻⁵ mol·L⁻¹, the stronger interaction of Triton X-100 with Hb was predominant. Hb was unfolded and denaturized. A little Triton X-100 can protect Hb from the effects of ribavirin.