Reversible Aggregation of Chlorophyll Derivative Induced by Phase Transition of Lipid.

Controlling the supramolecular organization of pigment molecules will provide innovative materials that exhibit variable optical properties. In nature, photosynthetic systems employ chlorophyllous supramolecules in which each pigment molecule is suitably organized in proteins, and their properties are adequately optimized by changing the structures of the surrounding amino acid residues. Here, we report a strategy for varying the aggregation behavior of a chlorophyll derivative by using a phase-transition phenomenon of lipid bilayers. Methyl pyropheophorbide a (MPP) was employed as a chlorophyllous pigment in our artificial system, and synthetic phosphatidylcholines with saturated acyl chain(s) were also used. The MPP molecules successfully accumulated within the lipid bilayer of liposomes without changing the vesicular structure. When the lipid bilayer was in a gel form (under the phase-transition temperature, Tm), the embedded MPP aggregated to yield a dimeric form showing red-shifted absorption bands and circular dichroism signals. When the solutions of MPP-containing liposomes were heated to higher temperatures than their Tm, MPP disaggregated to monomeric form as the absorption spectrum changed into its original fashion in dichloromethane. The reversible thermochromic (dis)aggregation of the MPP molecules had good cyclability. Additional careful examination of the phase transition in the MPP-lipid co-assemblies clarified that the critical temperatures of the MPP (dis)aggregation were in good agreement with the phase-transition temperatures of the pigment-containing bilayers. The reversible MPP aggregation in the lipid bilayers occurred in a wide range of temperatures (around 10-55 °C) by changing the length of the diacyl side chains of phospholipids. The reversible thermochromism of the chlorophyllous system was established by varying the nature of the surrounding lipid bilayer. This study can provide a useful strategy for making variable tetrapyrrolic aggregate systems induced by mild extrinsic stimuli.

Ultrafast dynamics of multi-exciton state coupled to coherent vibration in zinc chlorin aggregates for artificial photosynthesis.

Ultrafast vibronic dynamics induced by the interaction of the Frenkel exciton with the coherent molecular vibrations in a layer-structured zinc chlorin aggregates prepared for artificial photosynthesis have been studied by 7.1 fs real-time vibrational spectroscopy with multi-spectrum detection. The fast decay of 100 ± 5fs is ascribed to the relaxation from the higher multi-exciton state (MES) to the one-exciton state, and the slow one of 863 ± 70fs is assigned to the relaxation from Q-exciton state to the dark nonfluorescent charge-transfer (CT) state, respectively. In addition, the wavelength dependences of the exciton-vibration coupling strength are found to follow the zeroth derivative of the transient absorption spectra of the exciton. It could be explained in term of the transition dipole moment modulated by dynamic intensity borrowing between the B transition and the Q transition through the vibronic interactions.

Excitonic and vibrational coherence in artificial photosynthetic systems studied by negative-time ultrafast laser spectroscopy.

Quantum coherences between excitonic states are believed to have a substantial impact on excitation energy transfer in photosynthetic systems. Here, the excitonic and vibrational coherence relaxation dynamics of artificially synthetic chlorosomes are studied by a sub 7 fs negative-time-delay laser spectroscopy at room temperature. The results provide direct evidence for the quantum coherence of the excitonic dephasing time of 23 ± 1 fs at physiologically relevant temperatures, which is significant in the initial step of energy transfer in chlorosome or chlorosome-like photosynthetic systems. Meanwhile, coherent molecular vibrations in the excited state are also detected without the effect of wave-packet motion in the ground state, which shows that the excited state wave-packet motion contributes greatly to the vibrational modes of ∼150 and ∼1340 cm(-1) in artificial chlorosome systems.

Synthesis of chlorophyll-a derivatives methylated in the 3-vinyl group and their intrinsic site energy.

Wittig reaction of methyl pyropheophorbide-d possessing the 3-formyl group gave readily methyl pyropheophorbides-a bearing a variety of 3-alkenyl groups as semi-synthetic models of chlorophyll-a. The 3-substituents rotated around the C3-C3(1) bond from the coplanar conformation with the chlorin π-system, moving the redmost visible absorption maxima to a shorter wavelength. The model experiments showed that natural chlorophyll-a carrying the 3-vinyl group would take a similar rotamer to control its intrinsic site energy.

Assemblies of ionic zinc chlorins assisted by water-soluble polypeptides.

Artificial chlorophyll-peptide complexes were prepared by mixing ionic chlorophyll derivatives and oligopeptides in aqueous media containing a small amount of organic solvent. The pigment-peptide complexes provided chlorophyll assemblies showing a sharp red-shifted Qy absorption band concomitant with giant circular dichroism signals. The polypeptide-assisted assemblies of chlorophyllous pigments would afford a good model for photosynthetic apparatuses such as pigment-protein complexes of light-harvesting antennas.

Excitonic Relaxation and Coherent Vibrational Dynamics in Zinc Chlorin Aggregates for Artificial Photosynthetic Systems.

The excitonic relaxation and coherent vibrational dynamics in stairlike zinc chlorin aggregates prepared for mimicking chlorosome in nature have been studied simultaneously by 6.8 fs real-time vibrational laser spectroscopy. The relaxation from Q-exciton state to the dark nonfluorescent charge-transfer (CT) state is determined to be 850 ± 70 fs. The spectral distribution of the molecular vibrational amplitude has been discussed in terms of the difference in the equilibrium positions of potential curves between the ground state and the excited state. Since the displacement in the coordinate space from the potential minimum of the ground state to that of the excited states is small, coherent oscillations generated by the impulsive excitation are strongest where the slope of the excitonic resonance is largest. Consequently, the probe wavelength dependence of the amplitude modulation follows the first derivative of the excitonic resonance, and π phase jump has been observed. Excitonic transition energy modulation caused by the coherent molecular vibrations has also been studied, and the vibrational mode with a low frequency of 146 cm(-1) is found to play a dominating role in the transition energy shift effect.

Photoinduced cytotoxicity of a photochromic diarylethene via caspase cascade activation.

The photo-generated closed-ring isomer of bis(5-methyl-2-phenylthiazoyl)perfluorocyclopentene shows cytotoxicity to Madin-Darby canine kidney (MDCK) cells through a caspase cascade and induces apoptosis of cells.

Non-enzymatic conversion of chlorophyll-a into chlorophyll-d in vitro: a model oxidation pathway for chlorophyll-d biosynthesis.

Chlorophyll-a (Chl-a) was readily converted into Chl-d under mild conditions without any enzymes. Treatment of Chl-a dissolved in dry tetrahydrofuran (THF) with thiophenol and acetic acid at room temperature successfully produced Chl-d in 31% yield. During the acidic oxidation, removal of the central magnesium, pheophytinization, was sufficiently suppressed. This mild pathway can give insights into the yet unidentified Chl-d biosynthesis.

Magneto-chiral dichroism of artificial light-harvesting antenna.

We have demonstrated the presence of magneto-chiral dichroism (MChD) of chiral J-aggregates of zinc chlorins. To the best of our knowledge, this is the first observation of MChD in artificial light-harvesting antennas.

Synthetic pores with reactive signal amplifiers as artificial tongues.

The sensation of taste is mediated by activation or deactivation of transmembrane pores. Artificial stimulus-responsive pores are enormously appealing as sensor components because changes in their activity are readily detectable in many different ways. However, the detection of multiple components in complex matrices (such as foods) with one pore sensor has so far remained elusive because the specificity necessary for sensing a target compound in complex mixtures is incompatible with the broad applicability needed for the detection of multiple components. Here, we present synthetic pores that, like our tongues, can sense flavours in food and in addition make them visibly detectable. Differential sensing and pattern recognition are solutions based on empirical and biomimetic approaches. They have been explored with synthetic receptor arrays and electronic tongues. In contrast, our approach is non-empirical as it exploits reactive amplifiers that covalently capture elusive analytes after enzymatic signal generation and drag them into synthetic pores for blockage. Reactive amplification proved to be highly sensitive and adaptable to various analytes and pores. Moreover, it can be combined with reactive filtration for minimizing interference. The system was tested on real food samples for detection of sucrose, lactose, lactate, acetate, citrate and glutamate to demonstrate the feasibility of these synthetic pores as universal sensors.

A cost-effective method for the optical transduction of chemical reactions. Application to hyaluronidase inhibitor screening with polyarginine-counteranion complexes in lipid bilayers.

We report the systematic elaboration of a cost-effective, interference-minimized assay for the label-free optical transduction of chemical reactions. Recently, we have found that certain complexes formed by arginine-rich cell-penetrating peptides (CPPs) and amphiphilic counteranions can act as synergistic anion carriers in lipid bilayer membranes. Application of this discovery to rapid and reversible cytosolic CPP delivery has been described (Futaki, S.; et al. ACS Chem. Biol. 2006, 1, 299). Here, we report the complementary use of polyarginine (pR)-counteranion complexes as general optical transducers of chemical reactions. Counterion screening revealed dodecyl phosphate (DP) as an ideal pR activator. Carboxyfluorescein (CF)-loaded vesicles with a shelf life of 3.5 years served best for the detection of fluorogenic CF release by pR-DP complexes with the naked eye. Inactivation of pR-DP complexes by counterion exchange with hyaluronan (HA) caused no CF emission, while HA removal by hyaluronidase (HAase) did. pR-DP complexes were further compatible with the optical detection of HA immobilization on solid support as well as inhibitor screening for HAase (cromolyn, heparin) with and without substrate immobilization. Controls concerning binary ATP/ADP discrimination for naked-eye kinase detection are mentioned to delineate scope but also limitations of this simple and quite universal method.

Self-assembly of natural light-harvesting bacteriochlorophylls of green sulfur photosynthetic bacteria in silicate capsules as stable models of chlorosomes.

Naturally occurring bacteriochlorophyll(BChl)s-c, -d, and -e from green sulfur photosynthetic bacteria were self-assembled in an aqueous solution in the presence of octadecyltriethoxysilane and tetraethoxysilane, followed by polycondensation of the alkoxysilanes by incubation for 50 h at 25 degrees C. The resulting BChl self-assemblies in silicate capsules exhibited visible absorption and circular dichroism spectra similar to the corresponding natural light-harvesting systems (chlorosomes) of green sulfur bacteria. Dynamic light scattering measurements indicated that the silicate capsules had an average hydrodynamic diameter of several hundred nanometers. BChl self-aggregates in silicate capsules were significantly stable to a nonionic surfactant Triton X-100, which was apt to decompose the BChl aggregates to their monomeric form, compared with conventional micelle systems. BChls in silicate capsules were more tolerant to demetalation of the central magnesium under acidic conditions than the natural systems.

Time-dependent self-assembly of 31-epimerically pure and mixed zinc methyl bacteriopheophorbides-d in an aqueous THF solution.

The self-aggregation process of 3(1)-epimerically pure and mixed zinc methyl bacteriopheophorbides-d (ZMBPhes-d) was examined by stopped-flow technique. A 33(v/v)% tetrahydrofuran (THF) - water solution of ZMBPhe-d was rapidly mixed with a 7(v/v)% THF - water solution to form a chlorosome-type aggregate with a red-shifted Qy band around 700 nm. We observed a rapid autocatalytic aggregation in a subsecond time scale. Aggregates of the 3(1)R epimer increased with a change in the Qy absorption maximum from 698 to 705 nm, suggesting that small aggregates formed as intermediate species. In addition, the rate of aggregation was dependent on the stereochemistry at the 3(1)-position of ZMBPhe-d; the 3(1)R epimer self-aggregated more rapidly than the 3(1)S epimer.

Coaggregate of amphiphilic zinc chlorins with synthetic surfactants in an aqueous medium to an artificial supramolecular light-harvesting system.

Aqueous assemblies of zinc chlorins possessing a nonionic (oligo)oxyethylene, a cationic quaternary ammonium or an anionic sulfonate group were prepared in the presence of a synthetic surfactant. The nonionic zinc chlorin formed aggregates when admixed with a nonionic surfactant such as Triton X-100 to give a highly ordered oligomeric J-aggregate similarly as natural bacteriochlorophyll-c or d does in a chlorosome. In addition, the coassemblies of the cationic zinc chlorin with an anionic surfactant and of the anionic zinc chlorin with a cationic surfactant gave large oligomers of these chlorophyllous pigments. The structures of hydrophilic groups in both the zinc chlorin and surfactant molecules controlled their aqueous coassemblies.

Self-assembly of synthetic zinc chlorins in a silicate micelle prepared by sol-gel process.

Zinc methyl 3-devinyl-3-hydroxymethyl-pyropheophorbide-a (1), a good model compound of light-harvesting pigments of green photosynthetic bacteria, formed self-aggregates in the presence of octadecyltriethoxysilane and tetraethoxysilane in an aqueous solution to exhibit visible absorption spectra similar to the natural antenna. Base-catalyzed cross-linked polymerization of the additive Si-ORs (R=ethyl and H) afforded the formation of a siloxane network (Si-O-Si) on the surface of the self-assemblies of 1. The resulting microcapsules were stable to tolerate the deaggregation to monomeric 1 by addition of surfactant Triton X-100 more largely than the corresponding micelles before polymerization.

Regio- and stereoisomeric control of the aggregation of zinc-chlorins possessing inverted interactive hydroxyl and carbonyl groups.

As models for a self-aggregative, naturally occurring magnesium-chlorin bacteriochlorophyll-d possessing 3(1)-secondary alcoholic hydroxyl and 13(1)-oxo groups, zinc-chlorins were synthesized with 3(1)-oxo and 13(1)-secondary (1) or tertiary hydroxyl groups (2). Compared to the monomers in a tetrahydrofuran solution, diastereomers 13(1)R-1R and 13(1)S-1S gave red-shifted absorption maxima (643 --> 674 nm in 1R and 708 nm in 1S) in 1 v/v% CH(2)Cl(2)-hexane solution, indicating their self-aggregation. Therefore, the positioning of the two groups at 3(1)/13(1) or 13(1)/3(1) on the N21-N23 molecular (Q(y)) axis is not necessarily important for the self-aggregation. The (1)H NMR and CD spectroscopic studies showed that the 674 nm absorbing species of 1R was characterized as a face-to-face "closed" dimer, while the 708 nm absorbing species of 1S was a large oligomer constructed with aggregation of head-to-tail "open" dimers. This diastereomeric control over the aggregation of 1R and 1S is more pronounced than that observed in the regioisomerically 3(1)-secondary alcoholic R/S-diastereomers 3R and 3S. The difference is ascribable to the conformational fixation of the 13(1)-hydroxyl group of the exo five-membered ring in 1. In contrast to self-aggregative 3(1)-tertiary alcoholic 4, both 13(1)-epimers of 13(1)-tertiary alcoholic 2 were monomeric even in nonpolar organic media: the additional 13(1)-methyl group (1 --> 2) drastically suppressed the self-aggregation due to the interference of the methyl group in intermolecular pi-pi interaction.