The intrinsic pKa values for phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine in monolayers deposited on mercury electrodes.

The intrinsic pKa values of the phosphate groups of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) and of the phosphate and carboxyl groups of phosphatidylserine (PS) in self-organized monolayers deposited on a hanging mercury drop electrode were determined by a novel procedure based on measurements of the differential capacity C of this lipid-coated electrode. In view of the Gouy-Chapman theory, plots of 1/C at constant bulk pH and variable KCl concentration against the reciprocal of the calculated diffuse-layer capacity Cd,0 at zero charge exhibit slopes that decrease from an almost unit value to vanishingly low values as the absolute value of the charge density on the lipid increases from zero to approximately 2 microC cm-2. The intrinsic pKa values so determined are 0.5 for PE and 0.8 for PC. The plots of 1/C against 1/Cd,0 for pure PS exhibit slopes that pass from zero to a maximum value and then back to zero as pH is varied from 7.5 to 3, indicating that the charge density of the lipid film passes from slight negative to slight positive values over this pH range. An explanation for this anomalous behavior, which is ascribed to the phosphate group of PS, is provided. Interdispersion of PS and PC molecules in the film decreases the "formal" pKa value of the latter group by about three orders of magnitude

Electrochemical modeling of electron and proton transfer to ubiquinone-10 in a self-assembled phospholipid monolayer.

Ubiquinone-10 (UQ) was incorporated at concentrations ranging from 0.5 to 2 mol% in a self-assembled monolayer of dioleoylphosphatidylcholine (DOPC) deposited on a mercury drop electrode, and its electroreduction to ubiquinol (UQH2) was investigated in phosphate and borate buffers over the pH range from 7 to 9.5 by a computerized chronocoulometric technique. The dependence of the applied potential for a constant value of the faradaic charge due to UQ reduction upon the electrolysis time t at constant pH and upon pH at constant t was examined on the basis of a general kinetion approach. This permitted us to conclude that the reduction of UQ to UQH2 in DOPC monolayers takes place via the reversible uptake of one electron with the formation of the semiubiquinone radical anion UQ.-, followed by the rate-determining protonation of this anion with UQH. formation; this neutral radical is more easily reduced than UQ, yielding the ubiquinol UQH2. In spite of the very low concentration of hydrogen ions as compared with that of the acidic component of the buffer, the only effective proton donor is the proton itself; this strongly suggests that the protonation step takes place inside the polar head region of the DOPC monolayer, which is only accessible to protons

Voltammetry of L-cysteine and 2-mercaptopyridine on a self-assembled phospholipid monolayer

The redox behaviour of 2-mercaptopyridine and the aminoacid L-cysteine was studied through a self-assembled monolayer of dioleoylphosphatidylcholine adsorbed on mercury by using cyclic voltammetry. 2-Mercaptopyridine penetrates into monolayer in the zone of stability of the phospholipid layer and shows an quasi-reversible behaviour while reversibility is observed in the absence of a monolayer. This fact is reflected in the occurrence of voltammetric peaks in the above mentioned region of potentials. Conversely, cysteine was found not to penetrate in the lipid layer as long as the latter behaves like a half-membrane. Voltammetric signal of cysteine was only obtained when the potential was scanned to values positive to -0.2 V. Beyond this potential the cyclic voltammograms show a series of anodic peaks, due to a rearrangement of the lipid film and to the formation of Hg(RS)2, followed by three cathodic peaks when the scan is reversal. The behaviour of these peaks was analized

A Peptide-tethered Lipid Bilayer on Mercury as a Biomimetic System

A novel spacer consisting of a hexapeptide molecule with a high tendency to form a 310-helical structure, which terminates with a sulfydryl group for anchoring to a metal, was tailored for use as a tethered hydrophilic spacer to be interposed between a metal support and a lipid bilayer. The thiol peptide has two triethylenoxy side chains that impart it a satisfactory hydrophilicity and are intended to keep the anchored thiol peptide chains sufficiently apart so as to accommodate water molecules and inorganic ions and to create a suitable environment for the incorporation of integral proteins. This thiol peptide was anchored to a hanging mercury drop electrode. The formation of a phospholipid bilayer on top of the self-assembled thiol peptide was carried out by a novel procedure which exploits the spontaneous tendency of a lipid film to form a bilayer when interposed between two hydrophilic phases. The resulting mercury-supported thiol peptide/lipid bilayer system was characterized by ac voltammetry with phase resolution, chronocoulometry, and impedance spectroscopy. The suitability of this tethered film as a biomembrane model was tested by incorporating ubiquinone-10 and valinomycin