ABSTRACT
We propose surface-enhanced optical-mid-infrared photothermal (MIP) microscopy using highly crystalline silver nanowires, acting as a Fabry-Perot resonator, and demonstrate its applicability to enhanced mid-infrared surface sensing of thin polymer layers as thin as 20 nm.
ABSTRACT
There is a great need for the analysis of the chemical composition, structure, functional groups, and interactions at polymer-metal interfaces in terms of adhesion, corrosion, and insulation. Although atomic force microscopy-based infrared (AFM-IR) spectroscopy can provide chemical analysis with nanoscale spatial resolution, it generally requires to thin a sample to be placed on a substrate that has low absorption of infrared light and high thermal conductivity, which is often difficult for samples that contain hard materials such as metals. This study demonstrates that the combination of AFM-IR with low-angle microtomy (LAM) sample preparation can analyze buried polymer-metal interfaces with higher spatial resolution than that with the conventional sample preparation of a thick vertical cross-section. In the LAM of a polymer layer on a metal substrate, the polymer layer is tapered to be thin in the vicinity of the interface, and thus, sample thinning is not required. An interface between an epoxyacrylate layer and copper wire in a flexible printed circuit cable was measured using this method. A carboxylate interphase layer with a thickness of â¼130 nm was clearly visualized at the interface, and its spectrum was obtained without any signal contamination from the neighboring epoxyacrylate, which was difficult to achieve on a thick vertical cross-section. The combination of AFM-IR with LAM is a simple and useful method for high-spatial-resolution chemical analysis of buried polymer-metal interfaces.
ABSTRACT
Organic matter in carbonaceous chondrites is distributed in fine-grained matrix. To understand pre- and postaccretion history of organic matter and its association with surrounding minerals, microscopic techniques are mandatory. Infrared (IR) spectroscopy is a useful technique, but the spatial resolution of IR is limited to a few micrometers, due to the diffraction limit. In this study, we applied the high spatial resolution IR imaging method to CM2 carbonaceous chondrites Murchison and Bells, which is based on an atomic force microscopy (AFM) with its tip detecting thermal expansion of a sample resulting from absorption of infrared radiation. We confirmed that this technique permits â¼30 nm spatial resolution organic analysis for the meteorite samples. The IR imaging results are consistent with the previously reported association of organic matter and phyllosilicates, but our results are at much higher spatial resolution. This observation of heterogeneous distributions of the functional groups of organic matter revealed its association with minerals at â¼30 nm spatial resolution in meteorite samples by IR spectroscopy.
ABSTRACT
It has been experimentally shown that the folding of apoplastocyanin (apoPC) accompanies a very large enthalpic loss [N. Baden et al., J. Chem. Phys. 127, 175103 (2007)]. This implies that an even larger entropic gain occurs in stabilizing the folded structure to overcome the enthalpic loss. Here, we calculate the water-entropy gain upon the folding of apoPC using the angle-dependent integral equation theory combined with the multipolar water model and the recently developed morphometric approach. It is demonstrated that the calculated value is in quantitatively good accord with the value estimated from the experimental data by accounting for the conformational-entropy loss. According to a prevailing view, the water adjacent to a hydrophobic group is unstable especially in terms of the rotational entropy and the folding is driven primarily by the release of such unfavorable water to the bulk through the burial of nonpolar side chains. We show, however, that the resultant entropic gain is too small to elucidate the experimental result. The great entropic gain observed is ascribed to the reduction in the restriction for the translational motion of water molecules in the whole system.
Subject(s)
Apoproteins/chemistry , Apoproteins/metabolism , Models, Molecular , Plastocyanin/chemistry , Plastocyanin/metabolism , Protein Folding , Absorption , Entropy , Protein Conformation , Surface Properties , Thermodynamics , Water/chemistryABSTRACT
A method of diffusion coefficient (D) measurement for proteins based on the pulsed laser-induced transient grating method using a photosensitive cross-linker was applied to the characterization of the pH denaturation process of holo- and apo-myoglobin (Mb) from the viewpoint of protein-water interaction. It was found that the pH denaturation curve monitored by D agrees quite well with that determined by the circular dichroism intensity for holo-Mb. This fact indicates that the changes in intermolecular interaction and the alpha-helix content occur simultaneously during the unfolding process. However, the pH dependence of D for apo-Mb was different from that of alpha-helix content. This different behavior can be explained in terms of the different denaturation steps for the secondary structure and the hydrogen bonding network of the intermediate species around pH 4; i.e., this intermediate is partially unfolded, but the hydrogen bonding network is dominantly an intramolecular one. Taking previously reported properties of this species into account, we conclude that water molecules are trapped in the hydrophobic core of the apo-Mb pH 4 intermediate. This fact suggests that the kinetic intermediate state of the protein folding process is a swollen state without water molecular exchange with the bulk phase.
Subject(s)
Azides/chemistry , Myoglobin/chemistry , Water/chemistry , Azides/radiation effects , Hydrogen Bonding , Hydrogen-Ion Concentration , Kinetics , Lasers , Molecular Structure , Myoglobin/radiation effects , Protein Denaturation , Protein FoldingABSTRACT
A new method to investigate the initial protein folding dynamics is developed based on a pulsed laser light triggering method and a unique transient grating method. The side chain of the cysteine residue of apoplastocyanin (apoPC) was site-specifically modified with a 4,5-dimethoxy-2-nitrobenzyl derivative, where the CD and 2D NMR spectra showed that the modified apoPC was unfolded. The substituent was cleaved with a rate of about 400 ns by photoirradiation, which was monitored by the disappearance of the absorption band at 355 nm and the increase in the transient grating signal. After a sufficient time from the photocleavage reaction, the CD and NMR spectra showed that the native beta-sheet structure was recovered. Protein folding dynamics was monitored in the time domain with the transient grating method from a viewpoint of the molecular volume change and the diffusion coefficient, both of which reflect the global structural change, including the protein-water interaction. The observed volume decrease of apoPC with a time scale of 270 micros is ascribed to the initial hydrophobic collapse. The increase in the diffusion coefficient (23 ms) is considered to indicate a change from an intermolecular to an intramolecular hydrogen bonding network. The initial folding process of apoPC is discussed based on these observations.
Subject(s)
Apoproteins/chemistry , Light , Plastocyanin/chemistry , Protein Conformation/radiation effects , Protein Folding , Protein Structure, Secondary/radiation effects , Circular Dichroism , Cysteine/chemistry , Diffusion , Hydrogen Bonding , Hydrophobic and Hydrophilic Interactions , Lasers , Magnetic Resonance Spectroscopy , Nitrobenzenes/chemistry , Water/chemistryABSTRACT
The diffusion coefficients (D) of poly-L-glutamic acid (PLG) at various pHs are investigated by the laser-induced transient-grating method with a new photoreactive probe molecule. The pH dependence of D is compared with that of the helical content of PLG measured by circular dichroism. It is found that the pH dependences of both quantities are very similar. Since the frictions of the translational diffusion of charged and protonated carboxyl groups are found to be similar each other, it is concluded that the conformation of the main polymer chain is the main factor in determining the diffusion process; in other words, the alpha-helix conformation makes the molecular diffusion faster. This result indicates that the conformational change of a protein can be detected by monitoring the diffusion coefficient.