J. Jay Leitch

In Situ PM-IRRAS Studies of an Archaea Analogue Thiolipid Assembled on a Au(111) Electrode Surface

Polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) has been applied to determine the conformation, orientation, and hydration of a monolayer of 2,3-di-O-phytanyl-sn-glycerol-1-tetraethylene glycol-dl-alpha-lipoic acid ester (DPTL) self-assembled at a gold electrode surface. This Archaea analogue thiolipid has been recently employed to build tethered lipid bilayers. By synthesizing DPT(d16)L, a DPTL molecule with a deuterium substituted tetraethylene glycol spacer, it was possible to differentiate the C-H stretch vibrations of the phytanyl chains from the tetraethylene glycol spacer and acquire the characteristic IR spectra for the chains, spacer, and lipoic acid headgroup separately. Our results show that the structure of the monolayer displays remarkable stability in a broad range of electrode potentials and that the phytanyl chains remain in a liquid crystalline state. The tetraethylene glycol chains are coiled, and the IR spectrum for this region shows that it is in the disordered state. The most significant result of this study is the information that in contrast to expectations the spacer region is poorly hydrated. Our results have implications for the design of a tethered lipid membrane based on this thiolipid.

Electrochemical and PM-IRRAS a glycolipid-containing biomimetic membrane prepared using Langmuir-Blodgett/Langmuir-Schaefer deposition.

Differential capacitance, chronocoulometry, and polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) measurements were used to characterize the structure and orientation of a DMPC + cholesterol + GM 1 (60:30:10 mol %) bilayer supported at a Au(111) electrode surface prepared using combined Langmuir-Blodgett/Langmuir-Schaefer (LB/LS) deposition. The electrochemical measurements indicate that the incorporation of ganglioside GM 1 into the membrane significantly improves the quality of the bilayer, reflected in the very low capacitance value of approximately 0.8 microF cm (-2). An analysis of the infrared data suggests that the incorporation of the glycolipid into the membrane changes both the orientation of the lipid acyl chains in the membrane and the hydration of the membrane, particularly with respect to the interfacial region of the lipids.

Quantitative SNIFTIRS studies of (bi)sulfate adsorption at the Pt(111) electrode surface

Subtractively normalized interfacial Fourier transform infrared reflection spectroscopy (SNIFTIRS) was applied to study (bi)sulfate adsorption on a Pt(111) surface in solutions of variable pH while maintaining a constant total bisulfate/sulfate ((bi)sulfate) concentration without the addition of an inert supporting electrolyte. The spectra were recorded for both the p- and s-polarizations of the IR radiation in order to differentiate between the IR bands of the (bi)sulfate species adsorbed on the electrode surface from those species located in the thin layer of electrolyte. The spectra recorded with p-polarized light consist of the IR bands from both the species adsorbed at the electrode surface and those present in the thin layer of electrolyte between the electrode surface and ZnSe window whereas the s-polarized spectra contain only the IR bands from the species located in the thin layer of electrolyte. A new procedure was developed to calculate the angle of incidence and thickness of the electrolyte between the Pt(111) electrode surface and the ZnSe IR transparent window. By combining these values with the knowledge of the optical constants for Pt, H(2)O and ZnSe, the mean square electric field strength (MSEFS) at the Pt(111) electrode surface and for thin layer of solution were accurately calculated. The spectra recorded using s-polarization were multiplied by the ratio of the average MSEFS for p- and s-polarizations and subtracted from the spectra recorded using p-polarization in order to remove the IR bands that arise from the species present within the thin layer cavity. In this manner, the resulting IR spectra contain only the IR bands for the anions adsorbed on the Pt(111) electrode surface. The spectra of adsorbed anions show little change with respect to the pH ranging from 1 to 5.6. This behavior indicates that the same species is predominantly adsorbed on the metal surface for this broad range of pH values and the results suggest that sulfate is the most likely candidate for this adsorbate.

Electrochemical and PM-IRRAS characterization of cholera toxin binding at a model biological membrane.

A mixed phospholipid-cholestrol bilayer, with cholera toxin B (CTB) units attached to the monosialotetrahexosylganglioside (GM1) binding sites in the distal leaflet, was deposited on a Au(111) electrode surface. Polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) measurements were used to characterize structural and orientational changes in this model biological membrane upon binding CTB and the application of the electrode potential. The data presented in this article show that binding cholera toxin to the membrane leads to an overall increase in the tilt angle of the fatty acid chains; however, the conformation of the bilayer remains relatively constant as indicated by the small decrease in the total number of gauche conformers of acyl tails. In addition, the bound toxin caused a significant decrease in the hydration of the ester group contained within the lipid bilayer. Furthermore, changes in the applied potential had a minimal effect on the overall structure of the membrane. In contrast, our results showed significant voltage-dependent changes in the average orientation of the protein α-helices that may correspond to the voltage-gated opening and closing of the central pore that resides within the B subunit of cholera toxin.

Infrared studies of the potential controlled adsorption of sodium dodecyl sulfate at the Au(111) electrode surface.

Quantitative subtractively normalized interfacial Fourier transform infrared reflection spectroscopy (SNIFTIRS) was used to determine the conformation and orientation of sodium dodecyl sulfate (SDS) molecules adsorbed at the single crystal Au(111) surface. The SDS molecules form a hemimicellar/hemicylindrical (phase I) structure for the range of potentials between -200 ≤ E < 450 mV and condensed (phase II) film for electrode potentials ≥500 mV vs Ag/AgCl. The SNIFTIRS measurements indicate that the alkyl chains within the two adsorbed states of SDS film are in the liquid-crystalline state rather than the gel state. However, the sulfate headgroup is in an oriented state in phase I and is disordered in phase II. The newly acquired SNIFTIR spectroscopy measurements were coupled with previous electrochemical, atomic force microscopy, and neutron reflectivity data to improve the current existing models of the SDS film adsorbed on the Au(111) surface. The IR data support the existence of a hemicylindrical film for SDS molecules adsorbed at the Au(111) surface in phase I and suggest that the structure of the condensed film in phase II can be more accurately modeled by a disordered bilayer.

Polarization modulation infrared reflection-absorption spectroscopy studies of the influence of perfluorinated compounds on the properties of a model biological membrane.

A combination of the Langmuir-Blodgett and Langmuir-Schaefer techniques has been used to build a 1,2-dimyristoyl- sn-glycero-3-phosphocholine (DMPC) bilayer at a Au(111) electrode surface with hydrogen-substituted acyl chains in the top leaflet (solution side) and deuterium-substituted acyl chains in the bottom leaflet (gold side). Polarization modulation infrared reflection-absorption spectroscopy was used to determine changes in the conformation and orientation of the acyl chains of DMPC caused by the incorporation of two selected perfluorinated compounds, perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS), into the top leaflet of the bilayer. The incorporation of perfluorinated compounds into the DMPC bilayer caused a broadening of the methylene peaks and a shift in the methylene band positions toward higher frequencies. In addition, the tilt angle of the acyl chains decreased in comparison to the tilt angle of a pure DMPC bilayer. The reported tilt angles were smaller upon insertion of PFOS ( approximately 24 degrees ) than in the presence of PFOA ( approximately 30 degrees ). Overall, the results show that the incorporation of the perfluorinated acids has an effect on the bilayer similar to that of cholesterol by increasing the membrane fluidity and thickness due to a decrease in the tilt angle of the acyl chains.

Quantitative SHINERS analysis of temporal changes in the passive layer at a gold electrode surface in a thiosulfate solution.

Shell-isolated gold nanoparticles (SHINs) were employed to record shell-isolated nanoparticle-enhanced Raman spectra (SHINERS) of a passive layer formed at a gold surface during gold leaching from thiosulfate solutions. The (3-aminopropyl)triethoxysilane (APTES) and a sodium silicate solution were used to coat gold nanoparticles with a protective silica layer. This protective silica layer prevented interactions between the thiosulfate electrolyte and the gold core of the SHINs when the SHINs-modified gold electrode was immersed into the thiosulfate lixiviant. The SHINERS spectra of the passive layer, formed from thiosulfate decomposition, contained bands indicative of hydrolyzed APTES. We have demonstrated how to exploit the presence of these APTES bands as an internal standard to compensate for fluctuations of the surface enhancement of the electric field of the photon. We have also developed a procedure that allows for removal of the interfering APTES bands from the SHINERS spectra. These methodological advancements have enabled us to identify the species forming the passive layer and to determine that the formation of elemental sulfur, cyclo-S8, and polymeric sulfur chains is responsible for inhibition of gold dissolution in oxygen rich thiosulfate solutions.

Measurements of the Potentials of Zero Free Charge and Zero Total Charge for 1-thio- ± bβ-D-glucose and DPTL Modified Au(111) Surface in Different Electrolyte Solutions

Abstract The immersion method was used to measure the potential of zero free charge (Epzfc) of a bare Au(111) electrode and the electrode modified by self-assembled monolayers (SAMs) of 1-thio-β-D-glucose (β-Tg) and 2,3-di-O-phytanyl-sn-glycerol-1-tetraethylene glycol-D,L-α-lipoic acid ester (DPTL). The measurements were performed in three electrolyte solutions: 0.1 M NaH2PO4, 0.1 MKClO4 and 0.1 M NaF. The Epzc of the Au(111) electrode has different values in these electrolyte solutions due to the specific adsorption of phosphate and fluoride anions on the Au(111) surface. In contrast, when the gold surface is covered by a SAM of β-Tg, the adsorption of anions is suppressed and similar values of Epzfc were measured in the three electrolytes. Additional chronocoulometric experiments were performed to determine the potentials of the zero total charge (Epztc) for the thiol covered electrode. Significant differences between numerical values of potentials of the zero free charge Epzfc and potentials of the zero total charge Epztc were observed.

SEIRAS Studies of Water Structure in a Sodium Dodecyl Sulfate Film Adsorbed at a Gold Electrode Surface

Surface-enhanced infrared reflection-absorption spectroscopy (SEIRAS) was used to investigate the structure of water that is incorporated within a film of sodium dodecyl sulfate (SDS) adsorbed at a thin gold nanoparticle film deposited onto a silicon substrate. Previous studies on a Au(111) electrode surface showed that SDS molecules form long-range ordered hemicylindrical hemimicelles (phase I) for potentials -0.2 ≤ E ≤ 0.45 V vs Ag/AgCl and a disordered bilayer (phase II) for potentials E ≥ 0.5 V vs Ag/AgCl. The SEIRA spectra demonstrated that the hemimicellar film is water-rich and contains both a network of hydrogen-bonded water and a disturbed network of hydrogen bonds consisting of monomeric and dimeric water in the hydrophobic region of the film. No network water was observed in phase II of the film. However, SEIRAS data showed that sulfate groups in the disordered bilayer are hydrated. The SEIRAS spectra of the film of SDS were compared to the previously measured spectra obtained using subtractively normalized interfacial Fourier transform IR spectroscopy (SNIFTIRS). The complementarity of the spectroscopic information obtained by these two techniques was demonstrated.

Quantitative Subtractively Normalized Interfacial Fourier Transform Infrared Reflection Spectroscopy Study of the Adsorption of Adenine on Au(111) Electrodes

Quantitative subtractively normalized interfacial Fourier transform infrared reflection spectroscopy (SNIFTIRS) was used to determine the molecular orientation and identify the metal-molecular interactions responsible for the adsorption of adenine from the bulk electrolyte solution onto the surface of the Au(111) electrode. The recorded p-polarized IR spectra of the adsorbed species were subtracted from the collected s-polarized IR spectra to remove the IR contributions of the vibrational bands of the desorbed molecules that are located within the thin layer cavity of the spectroelectrochemical cell. The intense IR band around 1640 cm(-1), which is assigned to the pyrimidine ring stretching vibrations of the C5-C6 and C6-N10 bonds, and the IR band at 1380 cm(-1), which results from a combination of the ring stretching vibration of the C5-C7 bond and the in-plane CH bending vibration, were selected for the quantitative analysis measurements. The transition dipoles of these bands were evaluated by DFT calculations. Their orientations differed by 85 ± 5°. The tilt angles of adsorbed adenine molecules were calculated from the intensity of these two vibrations at different potentials. The results indicate that the molecular plane is tilted at an angle of 40° with respect to the surface normal of the electrode and rotates by 16° around its normal axis with increasing electrode potential. This orientation results from the chemical interaction between the N10 and gold atoms coupled with the π-π parallel stacking interactions between the adjacent adsorbed molecules. Furthermore, the changes in the molecular plane rotation with the electric field suggests that the N1 atom of adenine must also participate in the interaction between the molecule and metal.