Search

Chronocoulometry was applied to determine charge in a monolayer of nucleolipid (1,2-dipalmitoyl-sn‑glycero-3-(cytidine diphosphate)) deposited at a gold electrode surface. The immersion method was used to measure the potential of zero charge of the interface (Epzci), which is a sum of charge on the monolayer of the nucleolipid and charge on the gold surface. Photon polarization infrared reflection absorption spectroscopy (PM-IRRAS) was used to determine formation of the Watson-Crick complex between terminal cytidine moiety of the nucleolipid and guanine (its complementary base) added to the solution. The combination of electrochemical and spectroscopic studies allowed one to demonstrate that the Watson-Crick complex is formed when the interface is positively charged. The potential applied to the electrode affects not only the complex formation but also orientation of the cytosine moiety. The complex is formed when the cytosine moiety is oriented assuming a small angle with respect to the electrode surface. Ministerio de Ciencia e Innovación CTQ2014-57515-C2-1-R, RED2018-102412-T Junta de Andalucía FQM202 Natural Sciences and Engineering Research Council of Canada (NSERC) RG-03958

Two 4-thio pseudo-glycolipids, a disulfide and a thiol, were synthesized as candidates to build tethered bilayer lipid membranes (tBLMs) on gold (111) surface. Monolayers of the disulfide and thiol were built on gold surfaces using the self-assembly and Langmuir-Blodgett (LB) transfer methods. Monolayers were prepared in several solvents and at various temperatures; their quality was assessed by differential capacitance. Best monolayers were achieved when the disulfide was self-assembled in ethanol. The quality of such monolayers was further improved by allowing the assembly to proceed under stirring of the solution at increased temperatures. The charge number per adsorbed molecule and surface concentration of disulfide on gold (111) surface were determined by chronocoulometry. A DPhPC/disulfide tBLM was then built by vesicle fusion of 1,2-diphytanyl-sn-glycero-3-phosphocholine (DPhPC) on top of the disulfide monolayer. The minimum capacitance of the gold electrode with tBLM (1.3 mF cm 2) was close to the value of a real cell membrane and the AFM force spectroscopy measurements showed that the DPhPC/disulfide tBLM had a thickness of 6.2 ± 0.6 nm consistent with the thickness expected for a bilayer. Finally, the potential of the tBLM to study transmembrane proteins was assessed by investigating the reconstitution of gramicidin A into the membrane by polarization modulation infrared absorption spectroscopy (PM-IRRAS). These results demonstrate that the disulfide is a good candidate to construct tBLMs on gold surface and to study transmembrane proteins. Natural Sciences and Engineering Research Council of Canada 2020-12-12

Unilamellar DMPC/DMPG vesicles in the absence and presence of alamethicin were fused onto the surface of a gold electrode modified with a 1-thio-β-d-glucose self-assembled monolayer. The resulting floating bilayer lipid membranes (fBLMs) were investigated using atomic force microscopy (AFM) and electrochemical impedance spectroscopy (EIS). A corrugated film structure was observed for the pure DMPC/DMPG fBLMs due to surface stress between the tightly packed lipids. These corrugations are removed by the addition of alamethicin suggesting the lipid-peptide interactions alleviate the overall surface stress creating a more uniform bilayer. Both DMPC/DMPG films in the absence and presence of alamethicin had thickness of 5.5 ± 0.9 nm demonstrating that alamethicin has a minimal effect on the overall bilayer thickness. However, a significant decrease in membrane resistivity was observed when alamethicin was inserted into the fBLM indicating that the peptides are forming ion conducting pores. A direct visualization of the alamethicin pores was obtained by molecular resolution AFM images revealing that the pores are not randomly dispersed throughout the bilayer, but instead form hexagonal aggregates. The diameter of an individual pore within the aggregates is equal to 2.3 ± 0.3 nm, which is consistent with the size of a hexameric pore predicted by molecular dynamics simulations. Additionally, the image revealed a broad size distribution of alamethicin aggregates, which explains the origin of multiple conductivity states observed for the incorporation of alamethicin into free standing bilayer lipid membranes. Natural Sciences and Engineering Research Council of Canada

The dissolution mechanism of nickel grown by the carbonyl process was investigated using laboratory Ni samples. These samples were purposely engineered to form an alternating lamellar structure of Ni layers grown under two limiting conditions. Three layers deposited from low nickel carbonyl gas concentrations (5%) at a decomposition temperature of 280 °C (LC/HT) were deposited between four layers formed from high nickel carbonyl gas concentrations (40%) at a decomposition temperature of 200 °C (HC/LT). These lamellar structures were used to simulate the two extremes found within an industrial decomposition chamber for nickel pellets grown using the Mond process. Cyclic voltammetry and fluctuations in the electrode potential were recorded during the 120 min constant current dissolution experiment. Complementary scanning electron microscopy (SEM) and white light interference microscopy (WLIM) measurements were used to monitor the changes in surface morphology, texture and roughness as a function of the dissolution time. The electrochemical measurements showed that the dissolution of the sample proceeded in the transpassive regime at overvoltages above 300 mV. Large oscillations were observed in the chronopotentiogram, which suggests that the passive layer breaks down and then regrows on the surface of the nickel samples. SEM and WLIM images revealed that pits with lacy coverings were formed during the initial stages of nickel dissolution. With increasing dissolution time, the LC/HT regions dissolved at a significantly faster rate than the HC/LT regions and the lacy covers were more readily removed causing the transition to an open pit dissolution mechanism. The formation of trenches at boundaries between the lamina was observed at longer dissolution times. The average depth and width of the trenches also increased as a function of dissolution time. The results from the laboratory-grown lamellar nickel samples in this study clearly show preferential dissolution at boundaries between the lamina and that the dissolution of the LC/HT regions proceeds faster than the dissolution of the HC/LT regions. Natural Sciences and Engineering Research Council (NSERC) of Canada and Vale Canada

Polarized attenuated total reflection (ATR-IR) spectroscopy and fluorescence microscopy techniques were used to characterize a 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) membrane supported on porous, cationic hydrogel beads. Fluorescence microscopy images showed that the DPhPC coated the external surface of the hydrogel scaffold. In addition, a fluorescence assay of the emission intensity of the Tb3+/dipicolinic acid complex demonstrated that the DPhPC coating acted as a barrier to Tb3+ efflux from the scaffolded vesicle and successfully sealed the porous hydrogel bead. Fluorescence quenching and ATR-IR spectroscopic measurements revealed that the lipid coating has a bilayer structure. The phytanoyl chains were found to exhibit significant trans-gauche isomerization, characteristic of the fluid liquid phase. However, no lipid lateral mobility was observed by fluorescence recovery after photobleaching (FRAP) measurements. The phosphocholine headgroup was found to be well hydrated and oriented such that the cationic choline group tucked in behind the anionic phosphate group, consistent with an electrostatic attraction between the cationic scaffold and zwitterionic lipid. The absence of lipid lateral mobility may be due to the strength of this attraction. the Natural Science and Engineering Research Council of Canada (NSERC)