Molecular responses of proteins at different interfacial environments detected by sum frequency generation vibrational spectroscopy

Sum frequency generation (SFG) vibrational spectroscopy has been applied to investigate molecular responses of bovine serum albumin (BSA) molecules adsorbed at different interfacial environments. Molecular level and in situ SFG studies demonstrate that albumin molecules have different adsorption behaviors when contact with fused silica, polystyrene, and poly(methyl methacrylate). Adsorbed albumin molecules exhibit different structural changes when exposed to different chemical environments, including air, water, and hydrophobic solvents. This paper provides direct molecular insight into protein responses to different interfacial environments.     

Broadband vibrational sum frequency generation spectroscopy of a liquid surface.

An important advance in surface science has been the evolution of sum frequency generation to the application of studying surface structure and chemistry of liquid surfaces at the molecular-level by probing the vibrational signatures of surface molecules. Recently, broad-bandwidth sum frequency generation (BBSFG) spectroscopy has become an important tool for investigating gas-solid interfaces. BBSFG spectroscopy allows, theoretically, a surface sum frequency spectrum to be acquired within one pulse of the laser. In this paper, the viability of BBSFG to study inherently small nonlinear response interfaces and the time-resolving capability of this surface-selective technology are demonstrated. Presented here are the first published accounts of spectra from a liquid surface utilizing the broad-bandwidth sum frequency technology with acquisition times as low as 500 milliseconds.     

Molecular level studies of polymer behaviors at the water interface using sum frequency generation vibrational spectroscopy

Industrial plastics, biomedical polymers and numerous other polymeric systems are contacted with water for everyday functions and after disposal. Probing the interfacial molecular interactions between widely used polymers and water yields valuable information that can be extrapolated to macroscopic polymer/water interfacial behaviors so scientists can better understand polymer bio-compatibility, hygroscopic tendencies and improve upon beneficial polymer behavior in water. There is an ongoing concerted effort to elucidate the molecular level behaviors of polymers in water by using sum frequency generation vibrational spectroscopy (SFG). SFG stands out for its utility in probing buried interfaces in situ and in real time without disrupting interfacial chemistry. Included in this review are SFG water interfacial studies performed on poly(methacrylate) and (acrylate)s, poly(dimethyl siloxane)s, poly(ethylene glycol)s, poly(electrolyte)s and other polymer types. The driving forces behind common water/polymer interfacial molecular features will be discussed as well as unique molecular reorientation phenomena and resulting macroscopic behaviors from microscopic polymer rearrangement. © 2012 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys, 2013     

Detection and spectral analysis of trifluoromethyl groups at a surface by sum frequency generation vibrational spectroscopy

Sum frequency generation (SFG) vibrational spectroscopy was used to detect the presence of trifluoromethyl groups on the surface of 4-(trifluoromethyl)benzyl alcohol (TFMBA) in air. Supplementary data from infrared and Raman spectra were correlated to ab initio calculations by use of density functional theory (DFT) for TFMBA and three related compounds to reliably assign vibrational modes to the spectra. It was shown that strongly ordered CF3 groups dominate the surface of the TFMBA, and the vibrational modes of this functional group are strongly coupled to the benzene ring of the benzyl alcohol. This coupling, along with the SFG activity of the CF3 group, is removed with the insertion of an oxygen atom between the CF3 group and the benzene ring.     

Orientational motions of vibrational chromophores in molecules at the air/water interface with time-resolved sum frequency generation

The first time-resolved experiments in which interfacial molecules are pumped to excited electronic states and probed by vibrational sum frequency generation (SFG) are reported. This method was used to measure the out-of-plane rotation dynamics, i.e. time dependent changes in the polar angle, of a vibrational chromophore of an interfacial molecule. The chromophore is the carbonyl group, the rotation observed is that of the -C=O bond axis, with respect to the interfacial normal, and the interfacial molecule is coumarin 314 (C314) at the air/water interface. The orientational relaxation time was found to be 220+/-20 ps, which is much faster than the orientational relaxation time of the permanent dipole moment axis of C314 at the same interface, as obtained from pump-second harmonic probe experiments. Possible effects on the rotation of the -C=O bond axis due to the carbonyl group hydrogen bonding with interfacial water are discussed. From the measured equilibrium orientation of the permanent dipole moment axis and the carbonyl axis, and knowledge of their relative orientation in the molecule, the absolute orientation of C314 at the air/water interface is obtained.     

DPPC Langmuir monolayer at the air-water interface: probing the tail and head groups by vibrational sum frequency generation spectroscopy

Dipalmitoylphosphatidylcholine (DPPC) is the predominant lipid component in lung surfactant. In this study, the Langmuir monolayer of deuterated dipalmitoylphosphatidylcholine (DPPC-d62) in the liquid-expanded (LE) phase and the liquid-condensed (LC) phase has been investigated at the air-water interface with broad bandwidth sum frequency generation (BBSFG) spectroscopy combined with a Langmuir film balance. Four moieties of the DPPC molecule are probed by BBSFG: the terminal methyl (CD3) groups of the tails, the methylene (CD2) groups of the tails, the choline methyls (CH3) in the headgroup, and the phosphate in the headgroup. BBSFG spectra of the four DPPC moieties provide information about chain conformation, chain orientation, headgroup orientation, and headgroup hydration. These results provide a comprehensive picture of the DPPC phase behavior at the air-water interface. In the LE phase, the DPPC hydrocarbon chains are conformationally disordered with a significant number of gauche configurations. In the LC phase, the hydrocarbon chains are in an all-trans conformation and are tilted from the surface normal by 25 degrees. In addition, the orientations of the tail terminal methyl groups are found to remain nearly unchanged with the variation of surface area. Qualitative analysis of the BBSFG spectra of the choline methyl groups suggests that these methyl groups are tilted but lie somewhat parallel to the surface plane in both the LE and LC phases. The dehydration of the phosphate headgroup due to the LE-LC phase transition is observed through the frequency blue shift of the phosphate symmetric stretch in the fingerprint region. In addition, implications for lung surfactant function from this work are discussed.     

Ultrafast vibrational dynamics of water at a charged interface revealed by two-dimensional heterodyne-detected vibrational sum frequency generation

Two-dimensional heterodyne-detected vibrational sum frequency generation (2D HD-VSFG) spectroscopy is performed for an aqueous interface for the first time. The 2D HD-VSFG spectra in the OH stretch region are obtained from a positively charged surfactant∕water interface with isotopically diluted water (HOD∕D(2)O) to reveal the femtosecond vibrational dynamics of water at the charged interface. The 2D HD-VSFG spectrum is diagonally elongated immediately after photoexcitation, clearly demonstrating inhomogeneity in the interfacial water. This elongation almost disappears at 300 fs owing to the spectral diffusion. Interestingly, the 2D HD-VSFG spectrum at the 0 fs shows an oppositely asymmetric shape to the corresponding 2D IR spectrum in bulk water: The bandwidth of the bleach signal gets narrower when the pump wavenumber becomes higher. This suggests that the dynamics and mechanism of the hydrogen bond rearrangement at the charged interface are significantly different from those in bulk water.     

Sum frequency vibrational spectroscopy of leucine molecules adsorbed at air-water interface

Sum frequency vibrational spectroscopy was used to study adsorption of leucine molecules at air-water interface from solutions with different concentrations and pH values. The surface density and the orientation of the isopropyl head group of the adsorbed leucine molecules could be deduced from the measurements. It was found that the orientation depends on the surface density, but only weakly on bulk pH value at the saturated surface density. The vibrational spectra of the interfacial water molecules appeared to be strongly affected by the charge state of the adsorbed leucine molecules. Enhancement and inversion of polar orientation of interfacial water molecules by surface charges or field controllable by the bulk pH value were observed.     

Piperidine adsorption on hydrated alpha-alumina (0001) surface studied by vibrational sum frequency generation spectroscopy

The adsorption of piperidine vapor on the hydrated alumina (alpha-Al2O3, corundum) (0001) surface was investigated using vibrational broad bandwidth and scanning sum frequency generation (SFG) spectroscopy. The interfacial vibrational signature in the C-H stretching region of piperidine at the alumina (0001) surface is shown to be a sensitive spectroscopic probe revealing the adsorption mechanism. The neat piperidine surface, aqueous piperidine surface, and aqueous piperidium chloride surface were also investigated in the C-H stretching region by SFG to establish vibrational reference frequencies. After piperidine adsorption, piperidine vapor was removed and piperidine was found to be chemisorbed onto the alumina (0001) surface through protonation by surface hydroxyl groups. The O-H stretching region of the alumina surface before and after piperidine adsorption was also investigated, and the results revealed the decrease of the surface number density of alumina surface hydroxyl groups.     

Measuring polymer surface ordering differences in air and water by sum frequency generation vibrational spectroscopy

Molecular structures of poly(n-butyl methacrylate) (PBMA) at the PBMA/air and PBMA/water interfaces have been studied by sum frequency generation (SFG) vibrational spectroscopy. PBMA surfaces in both air and water are dominated by the methyl groups of the ester side chains. The average orientation and orientation distribution of these methyl groups at the PBMA/air and PBMA/water interfaces are different, indicating that surface restructuring occurs when the PBMA sample contacts water. Analysis shows that the orientation distribution of side chain methyl groups on the PBMA surface is narrower in water than that in air, indicating that the PBMA surface can be more ordered in water. To our knowledge, this is the first time that quantitative comparisons between molecular surface structures of polymers in air and in water have been made. Two assumptions on the orientation distribution function, including a Gaussian distribution and a formula based on the maximum entropy approach, are used in the analysis. It has been found that the orientation angle distribution function deduced by the Gaussian distribution and the maximum entropy distribution are quite similar, showing that the Gaussian distribution is a good approximation for the angle distribution. The effect of experimental error on the deduced orientational distribution is also discussed.     

Non-linear vibrational sum frequency spectroscopy of atmospherically relevant molecules at aqueous solution surfaces

Surface vibrational sum frequency spectroscopy has been shown to be a powerful surface probe of molecules adsorbed at solid and liquid surfaces. Studies described herein apply this method to studying heterogeneous air/aqueous solution interfaces to understand surface adsorption and structure of several solute molecules adsorbed at aqueous surfaces. The molecules examined at aqueous solution surfaces include Dimethyl sulfoxide (DMSO), methane sulfonic acid (MSA) and acetone. These results reveal that small soluble molecules such as these organize in different ways at the surface of aqueous solutions. This surface organization has implications for atmospheric chemical processes since adsorption at the surface of atmospheric aerosols affects bulk chemical concentrations.     

Molecular interactions of proteins and peptides at interfaces studied by sum frequency generation vibrational spectroscopy

Interfacial peptides and proteins are critical in many biological processes and thus are of interest to various research fields. To study these processes, surface sensitive techniques are required to completely describe different interfacial interactions intrinsic to many complicated processes. Sum frequency generation (SFG) spectroscopy has been developed into a powerful tool to investigate these interactions and mechanisms of a variety of interfacial peptides and proteins. It has been shown that SFG has intrinsic surface sensitivity and the ability to acquire conformation, orientation, and ordering information about these systems. This paper reviews recent studies on peptide/protein-substrate interactions, peptide/protein-membrane interactions, and protein complexes at interfaces and demonstrates the ability of SFG on unveiling the molecular pictures of complicated interfacial biological processes.     

Membrane orientation of MSI-78 measured by sum frequency generation vibrational spectroscopy

Antimicrobial peptides (AMPs) selectively disrupt bacterial cell membranes to kill bacteria whereas they either do not or weakly interact with mammalian cells. The orientations of AMPs in lipid bilayers mimicking bacterial and mammalian cell membranes are related to their antimicrobial activity and selectivity. To understand the role of AMP-lipid interactions in the functional properties of AMPs better, we determined the membrane orientation of an AMP (MSI-78 or pexiganan) in various model membranes using sum frequency generation (SFG) vibrational spectroscopy. A solid-supported single 1,2-dipalmitoyl-an-glycero-3-[phospho-rac-(1-glycerol)] (DPPG) bilayer or 1-palmitoyl-2-oleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (POPG) bilayer was used as a model bacterial cell membrane. A supported 1,2-dipalmitoyl-an-glycero-3-phosphocholine (DPPC) bilayer or a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayer was used as a model mammalian cell membrane. Our SFG results indicate that the helical MSI-78 molecules are associated with the bilayer surface with ～70° deviation from the bilayer normal in the negatively charged gel-phase DPPG bilayer at 400 nM peptide concentration. However, when the concentration was increased to 600 nM, MSI-78 molecules changed their orientation to make a 25° tilt from the lipid bilayer normal whereas multiple orientations were observed for an even higher peptide concentration in agreement with toroidal-type pore formation as reported in a previous solid-state NMR study. In contrary, no interaction between MSI-78 and a zwitterionic DPPC bilayer was observed even at a much higher peptide concentration (～12,000 nM). These results demonstrate that SFG can provide insights into the antibacterial activity and selectivity of MSI-78. Interestingly, the peptide exhibits a concentration-dependent membrane orientation in the lamellar-phase POPG bilayer and was also found to induce toroidal-type pore formation. The deduced lipid flip-flop from SFG signals observed from lipids also supports MSI-78-induced toroidal-type pore formation.     

Detection of tethered biocide moiety segregation to silicone surface using sum frequency generation vibrational spectroscopy

Polymer surface properties are controlled by the molecular surface structures. Sum frequency generation (SFG) vibrational spectroscopy has been demonstrated to be a powerful technique to study polymer surface structures at the molecular level in different chemical environments. In this research, SFG has been used to study the surface segregation of biocide moieties derived from triclosan (TCS) and tetradecyldimethyl (3-trimethoxysilylpropyl) ammonium chloride (C-14 QAS) that have been covalently bound to a poly(dimethylsiloxane) (PDMS) matrix. PDMS materials are being developed as coatings to control biofouling. This SFG study indicated that TCS-moieties segregate to the surface when the bulk concentration of TCS-moieties exceeds 8.75% by weight. Surface segregation of C-14 QAS moieties was detected after 5% by weight incorporation into a PDMS matrix. SFG results were found to correlate well with antifouling activity, providing a molecular interpretation of such results. This research showed that SFG can aid in the development of coatings for controlling biofouling by elucidating the chemical structure of the coating surface.     

Recent progress in theoretical analysis of vibrational sum frequency generation spectroscopy

This article summarizes the computational analysis of the vibrational sum frequency generation (SFG) spectroscopy with molecular dynamics simulation. The analysis allows direct comparison of experimental SFG spectra and microscopic interface structure obtained by molecular simulation, and thereby obviates empirical fitting procedures of the observed spectra. In the theoretical formulation, the frequency-dependent nonlinear susceptibility of an interface is calculated in two ways, based on the energy representation and time-dependent representation. The application to aqueous interfaces revealed a number of new insights into the local structure of electrolyte interfaces and the interpretation of SFG spectroscopy.     

Studying nanoparticle-induced structural changes within fatty acid multilayer films using sum frequency generation vibrational spectroscopy

The nonlinear optical technique of sum frequency generation (SFG) vibrational spectroscopy has been used for the first time to study CdS nanoparticle/arachidic acid multilayer structures. Using a combination of per-deuterated and per-protonated arachidic acid, it is possible to study individual layers anywhere within the film, buried or on the surface. Before reaction with H2S all layers are highly ordered, but after the reaction the layers become highly disordered, except for the surface layer, which remains well ordered. This sheds new light on the structure and stability of these films and shows that SFG can provide unique structural information.     

Chain conformation of poly(dimethyl siloxane) at the air/water interface by sum frequency generation

This is to report a study of chain conformation of poly(dimethylsiloxane) (PDMS) in spread monolayers at the air/water interface (A/W) with the aid of vibrational sum frequency spectroscopy (VSFS). We find that methyl groups of PDMS chains at the interface are completely disordered in the dilute regime of the surface density. At higher surface densities, however, the two methyl groups on the repeating unit point into the air asymmetrically; one points more normal to the interface, whereas the other lies more parallel to the interface. In the first collapsed regime, where the surface pressure of the PDMS monolayer reaches a plateau value of 8.7 mN/m, the signal intensity at 2915 cm (-1), assigned to the symmetric vibrational frequency of the methyl groups, is found independent of the surface density. On the basis of this finding, we propose that PDMS chains, in the first collapse regime at the A/W, form asymmetric layers. Thus, our proposal lends support to earlier works by Langevin's group to refute a widely speculated helix model that was based on energy minimization in the crystalline state of PDMS. In short, the energy consideration in the bulk crystalline state does not provide meaningful guidance as to the chain conformation of the monolayer at the A/W.     

Adsorption of SDS and PEG on calcium fluoride studied by sum frequency generation vibrational spectroscopy

The adsorption of sodium dodecyl sulfate (SDS) from aqueous solution onto a calcium fluoride substrate (CaF(2)), in the presence of polyethylene glycol (PEG) of different molecular weights, has been investigated using the interface specific nonlinear optical technique of sum frequency generation (SFG) vibrational spectroscopy. Spectra of adsorbed SDS (in the C-H stretching region) were recorded at the surface of a CaF(2) prism in contact with SDS solutions at concentrations up to the cmc (8 mM) of the pure surfactant and in contact with binary solutions containing SDS and PEG with molecular weights from 400 to 12 000. In contrast with SFG spectra from the same combinations of surfactant and polymer on a hydrophobic surface, there was no evidence of spectra arising from the actual polymer adsorbed on CaF(2) at any polymer molecular weight either in the absence or presence of surfactant. However, there was indirect evidence for the presence of adsorbed polymer from changes in the SDS SFG spectra in the presence of polymer compared with spectra when the polymer was absent. The SFG spectra of SDS at 0.8 mM were closely similar to each other at all polymer molecular weights and different from the spectra in the absence of the polymer. The spectral differences between the polymer present and polymer absent was much smaller when the solution concentration of surfactant was 8 mM.     

Potential-dependent vibrational spectroscopy of solvent molecules at the Pt(111) electrode in a water/acetonitrile mixture studied by sum frequency generation

Sum frequency generation (SFG) vibrational spectra of D(2)O and/or acetonitrile (CH(3)CN) on a Pt(111) single-crystal electrode were obtained as a function of applied potential in a 5 mol % water/acetonitrile mixed solvent with different 0.1 molar MSO(3)CF(3) salts (M = H(+), Li(+), Na(+), K(+), and Cs(+)). The results provide a very specific model for the composition of the inner Helmholtz layer as a function of potential and surface charge. Acetonitrile dominates the inner layer with the CN group directed toward the metal at potentials where the metal has a positive charge. As the surface becomes negatively charged, the acetonitrile orientation flips 180 degrees, with the CH(3) group pointing toward the surface. At even more negative surface charge, D(2)O displaces acetonitrile from the inner layer and is the predominant molecule on the surface. Here water is present as an oriented molecule with the oxygen end pointing toward the metal. The potential (and surface charge) where water is the dominant molecule in the inner Helmholtz layer is determined by the solvation energy of the cation.     

Monolayer vibrational spectroscopy by infrared-visible sum generation at metal and semiconductor surfaces

IR-visible sum generation spectroscopy, an interface-selective probe of molecular vibrations, is used to obtain vibrational spectra of molecular monolayers on metal and semiconductor surfaces. The spectra obey electric dipole selection rules: vibrational modes must be both Raman and infrared active to show sum frequency resonances. The orientation of molecules at the interface can be determined by interference between the resonant molecular signal and a substrate background signal. Sum generation is also observed at a buried interface in the absence of a dielectric discontinuity, suggesting uses at buried molecular structures such as polymer-polymer interfaces.