Raman spectrometer with vertical flow method for solutions containing organic solvents.

The vertical flow (VF) method improves generation and collection efficiency in Raman spectroscopy. It enhances all Raman signals, including undesired signals of organic solvents having a considerably large Raman cross section. We constructed a Raman spectrometer using the VF method to overcome this drawback and introduced a spatial line rejection mask to eliminate unnecessary bands.

ROS-Responsive Methionine-Containing Amphiphilic Peptides Impart Enzyme-Triggered Phase Transition and Antioxidant Cell Protection.

Reactive oxygen species (ROS) are produced by cellular activities, such as metabolism and immune response, and play important roles in cell signaling and homeostasis. However, overproduced ROS causes irreversible damage to nucleic acids and membrane lipids, supporting genetic mutations and enhancing the effects of aging. Cells defend themselves against ROS using antioxidant systems based on redox-active sulfur and transition metals.

Origin of unique hyper-Raman signals of trifluoroethanol.

We report the hyper-Raman (HR) spectrum of trifluoroethanol, excited with 532 nm light, in an aqueous solution at basic pH. The HR spectrum exhibits a distinct spectral pattern that diverges entirely from the infrared and Raman spectra of trifluoroethanol. This observed unique HR signal was attributed to the products of photoinduced radical reactions in the aqueous solution.

Determination of sugar content in honey using LC-Raman and programmable pump-Raman methods.

We combined (i) liquid chromatography and Raman spectrometry (LC-Raman) and (ii) programmable pump and Raman spectrometry (PP-Raman) to separate and identify compounds in a mixture. These techniques were applied to conduct a quantitative analysis of the sugars in honey. The spectral and temporal axes of the LC-Raman data were analyzed using the MCR-ALS analysis procedure, which enabled the separation and identification of four sugars (glucose, fructose, sucrose, and trehalose).

ROS-Triggered Gel-Sol Transition and Kinetics-Controlled Cargo Release by Methionine-Containing Peptides.

The gel-sol transition of self-assembling peptides is a useful switch for environment-dependent drug release. For their applications, kinetics control of the responses is important for matching the velocity of release to the target biological events. Here we demonstrate the chemical control of redox-triggered gel-sol transition kinetics of self-assembling peptides by altering the amino acid sequence.

Resonance Hyper-Raman Spectroscopy of Nucleotides and Polynucleotides.

We applied 532 nm-excited two-photon resonance hyper-Raman (RHR) spectroscopy to nucleotides (dA, dG, dT, and dC) to obtain fundamental knowledge about their spectral patterns. The RHR spectrum of each nucleotide exhibited various modes of the purine and pyrimidine rings, showing the ability to acquire the structural information on the chromophore. The band positions of the RHR spectrum and the 266 nm-excited one-photon UV-resonance Raman (UVRR) spectrum were identical, while the intensity patterns differed.

Efficient protein incorporation and release by a jigsaw-shaped self-assembling peptide hydrogel for injured brain regeneration.

During injured tissue regeneration, the extracellular matrix plays a key role in controlling and coordinating various cellular events by binding and releasing secreted proteins in addition to promoting cell adhesion. Herein, we develop a cell-adhesive fiber-forming peptide that mimics the jigsaw-shaped hydrophobic surface in the dovetail-packing motif of glycophorin A as an artificial extracellular matrix for regenerative therapy. We show that the jigsaw-shaped self-assembling peptide forms several-micrometer-long supramolecular nanofibers through a helix-to-strand transition to afford a hydrogel under physiological conditions and disperses homogeneously in the hydrogel.

Regulation of Cell Volume by Nanosecond Pulsed Electric Fields.

Stimulation of cells by nanosecond pulsed electric fields (nsPEFs) has attracted attention as a technology for medical applications such as cancer treatment. nsPEFs have been shown to affect intracellular environments without significant damage to cell membranes; however, the mechanism underlying the effect of nsPEFs on cells remains unclear. In this study, we constructed electrodes for applying nsPEFs and analyzed the change in volume of a single cell due to nsPEFs using fluorescence and Raman microscopy.

Hydrogel-Stiffening and Non-Cell Adhesive Properties of Amphiphilic Peptides with Central Alkylene Chains.

Invited for the cover of this issue is the group of Takahiro Muraoka at Tokyo University of Agriculture and Technology and collaborators. The image depicts nanofiber formation of an amphiphilic peptide with a central alkylene chain that shows non-cell adhesive properties. Read the full text of the article at 10.

Hydrogel-Stiffening and Non-Cell Adhesive Properties of Amphiphilic Peptides with Central Alkylene Chains.

Amphiphilic peptides bearing terminal alkyl tails form supramolecular nanofibers that are increasingly used as biomaterials with multiple functionalities. Insertion of alkylene chains in peptides can be designed as another type of amphiphilic peptide, yet the influence of the internal alkylene chains on self-assembly and biological properties remains poorly defined. Unlike the terminal alkyl tails, the internal alkylene chains can affect not only the hydrophobicity but also the flexibility and packing of the peptides.

pH-controlled stacking direction of the β-strands in peptide fibrils.

Peptides provide a framework for generating functional biopolymers. In this study, the pH-dependent structural changes in the 21-29 fragment peptide of β-microglobulin (βm) during self-aggregation, i.e.

Online Liquid Chromatography-Raman Spectroscopy Using the Vertical Flow Method.

Liquid chromatography and Raman spectroscopy (LC-Raman system) were combined and developed with the aid of the vertical flow method that enhances the Raman signal intensity. The LC-Raman system enabled the online acquisition of the nonresonance Raman spectrum of LC eluates. We employed singular value decomposition (SVD) and subsequent reconstruction of the components for the analysis of two-dimensional (temporal and spectral) data.

Linear, mixed-valent homocatenated tri-tin complexes featuring Sn-Sn bonds.

A series of tri-tin complexes (LPhSn)3X with triple-decker structures (LPh = 2,5-di(o-pyridyl)-3,4-diphenylpyrrolate; X = Cl, AlCl4, OTf, and PF6) was synthesized by reducing LPhSnCl with LiBsBu3H and subsequent reactions. Structural characterization of (LPhSn)3Cl revealed a Sn-Sn-Sn core, and DFT calculations suggest that its HOMO is primarily σ-bonding along the tri-tin framework. (LPhSn)3Cl reacts with W(CO)5THF to afford (LPhSn)2(W(CO)5)2 and LPhSnCl, implying that (LPhSn)3Cl may exhibit dynamic behavior in solution.

Time-resolved FTIR study on the structural switching of human galectin-1 by light-induced disulfide bond formation.

Disulfide bonds play a fundamental role in controlling the tertiary structure of proteins; the formation or cleavage of some disulfide bonds can switch the structures and/or functions of proteins. Human galectin-1 (hGal-1), which is a lectin protein, exemplifies how both structure and function are changed by disulfide bonds; the structure and sugar-binding ability of hGal-1 are altered by the formation and cleavage of its three intra-molecular disulfide bonds. In the present study, the dynamics of the structural change of hGal-1 by the formation of disulfide bonds were investigated by time-resolved FTIR spectroscopy combined with a modification in which its thiol groups (-SH) were replaced with S-nitrosylated groups (SNO).

Tautomer Structures in Ketose-Aldose Transformation of 1,3-Dihydroxyacetone Studied by Infrared Electroabsorption Spectroscopy.

The acyclic form of monosaccharides exists in a structural equilibrium, with aldose having the aldehyde group and ketose the ketone group (ketose-aldose equilibrium). A basic catalyst facilitates their transformation, which affects the chemical properties of the monosaccharide. In this study, we investigated the ketose-aldose transformation of 1,3-dihydroxyacetone (1,3-DHA), one of the simplest systems of the ketose-aldose equilibrium.

A Vertical Flow Method for Sensitive Raman Protein Measurement in Aqueous Solutions.

Raman scattering is intrinsically faint. Raman spectroscopy would be more valuable with improvements in the signal detection efficiency. To improve the signal detection efficiency, we propose a vertical flow method, which is a derivative of the liquid core optical fiber technique employed for sensitive Raman signal detection.

Preparation and photo-induced activities of water-soluble amyloid β-C complexes.

We have shown that fullerene (C) becomes soluble in water by mixing fullerene and amyloid β peptide (Aβ40) whose fibril structures are considered to be associated with Alzheimer's disease. The water-solubility of fullerene arises from the generation of a nanosized complex between fullerene and the monomer species of Aβ40 (Aβ40-C). The prepared Aβ40-C exhibits photo-induced activity with visible light to induce the inhibition of Aβ40 fibrillation and the cytotoxicity for cultured HeLa cells.

Directly Probing Intermolecular Structural Change of a Core Fragment of β-Microglobulin Amyloid Fibrils with Low-Frequency Raman Spectroscopy.

Amyloid fibrils, which are ordered aggregates of proteins or peptides, have attracted keen interest because their deposition causes serious human diseases. Despite many studies utilizing X-ray crystallography, solid-state NMR, and other methods, intermolecular interactions governing the fibril formation remain largely unclear. Here, we used low-frequency Raman (LFR) spectroscopy to investigate the intermolecular β-sheet structure of a core fragment of β-microglobulin amyloid fibrils, βm, in aqueous buffer solutions.

Citrullination and deamidation affect aggregation properties of amyloid β-proteins.

Citrullination and deamidation, which are aging-related posttranslational modifications, increase the number of negative charges on amyloid β-protein (Aβ) at neutral pH. We investigated the effects of these modifications on the fibrillation properties of Aβ. The Arg5→Cit modification of Aβ did not affect the fibrillation rate, and brought β-sheet structures unlike that in the Aβ fibril.

Generation of self-clusters of galectin-1 in the farnesyl-bound form.

Ras protein is involved in a signal transduction cascade in cell growth, and cluster formation of H-Ras and human galectin-1 (Gal-1) complex is considered to be crucial to achieve its physiological roles. It is considered that the complex is formed through interactions between Gal-1 and the farnesyl group (farnesyl-dependent model), post-translationally modified to the C-terminal Cys, of H-Ras. We investigated the role of farnesyl-bound Gal-1 in the cluster formation by analyzing the structure and properties of Gal-1 bound to farnesyl thiosalicylic acid (FTS), a competitive inhibitor of the binding of H-Ras to Gal-1.

Effects of N-Methylated Amyloid-β30-40 Peptides on the Fibrillation of Amyloid-β1-40.

Alzheimer's disease is a neurodegenerative disorder associated with amyloid-β (Aβ) fibrillation. N-Methylated amyloid-β peptides are potent inhibitors of amyloid-β fibrillation. We investigated the inhibitory effect of N-Methylated Aβ30-40 peptides on Aβ1-40 fibrillation.

[Structure analysis of disease-related proteins using vibrational spectroscopy].

Analyses of the structure and properties of identified pathogenic proteins are important for elucidating the molecular basis of diseases and in drug discovery research. Vibrational spectroscopy has advantages over other techniques in terms of sensitivity of detection of structural changes. Spectral analysis, however, is complicated because the spectrum involves a substantial amount of information.

Characterization of intermolecular structure of β(2)-microglobulin core fragments in amyloid fibrils by vacuum-ultraviolet circular dichroism spectroscopy and circular dichroism theory.

Intermolecular structures are important factors for understanding the conformational properties of amyloid fibrils. In this study, vacuum-ultraviolet circular dichroism (VUVCD) spectroscopy and circular dichroism (CD) theory were used for characterizing the intermolecular structures of β2-microglobulin (β2m) core fragments in the amyloid fibrils. The VUVCD spectra of β2m20-41, β2m21-31, and β2m21-29 fragments in the amyloid fibrils exhibited characteristic features, but they were affected not only by the backbone conformations but also by the aromatic side-chain conformations.

Enhancement of proton transport in an oriented polypeptide thin film.

Proton transport properties of a partially protonated poly(aspartic acid)/sodium polyaspartate (P-Asp) were investigated. A remarkable enhancement of proton conductivity has been achieved in the thin film. Proton conductivity of 60-nm-thick thin film prepared on MgO(100) substrate was 3.

Involvement of histidine residues in the pH-dependent β-galactoside binding activity of human galectin-1.

The pH dependence of the β-galactoside binding activity of human galectin-1 (hGal-1) was investigated by fluorescence spectroscopy using lactose as a ligand. The obtained binding constant Kb was 2.94 ± 0.