Lipid Membrane Interface Research Articles

Membrane microstructural templates for enzyme domain formation.

Soluble proteins can spontaneously self-organize into two-dimensional domains at membrane interfaces, given sufficient mobility and specificity to membrane-localized ligands. The authors' recent results studying interfacial domain formation of the membrane-active enzyme, phospholipase A2, indicate that lateral phase separation of heterogeneous membrane mixtures creates anionic templates of specific morphology onto which the enzyme deposits, forming large protein assemblies. Selective removal of membrane components (lysolipid or fatty acid) produces different enzyme interfacial responses and domain morphologies. This leads to the conclusion that complex chemical and physical interactions laterally in the lipid membrane interface as well as between bound protein molecules play a role in organizing protein structures.

A laser-T-jump study of the adsorption of dipolar molecules to planar lipid membranes. II. Phloretin and phloretin analogues.

Phloretin and structurally related neutral molecules adsorb to the interface of lipid membranes and modify the electric dipole potential of the membrane/water interface. The adsorption process has been studied using a laser-T-jump relaxation technique in combination with an analysis of nonactin mediated potassium transport (see part I, Awiszus and Stark 1988). Deviations from the Langmuir isotherm were observed for most of the substances. The discrepancies were most pronounced at large surface densities, whereas good agreement was found at low concentrations in many cases. The partition coefficient in the limit of low concentrations was compared with that of octanol/water bulk phases. No correlation was found. The individual values of the two partition coefficients differed by more than three orders of magnitude. The contribution, b, of a single adsorbed molecule to the dipole potential could not be predicted from the dipole moment, microL, of the molecule measured in the bulk phase. Different values of b were found at identical values of microL. The study shows the limitations of the use of bulk phase data to predict molecular properties in lipid membranes.

A laser-T-jump study of the adsorption of dipolar molecules to planar lipid membranes. I. 2,4-dichlorophenoxyacetic acid.

The adsorption of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) as well as of other dipolar molecules to the interface of artificial lipid membranes gives rise to a change of the dipole potential between the membrane interior and water. As a consequence of the adsorption of the neutral species, the conductance of planar membranes, observed in the presence of the macrocyclic ion carriers nonactin or valinomycin, may change by many orders of magnitude. Using this effect in combination with a laser-T-jump technique, the kinetics of the adsorption process were measured and were interpreted on the basis of a Langmuir-isotherm. A partition coefficient (at small concentrations) of beta HA = 4.7 x 10(-4) cm, a rate constant of desorption kHA greater than or equal to 100 s-1 and a maximum surface density ND = 7.7 x 10(13)/cm2 were found. The concentration at half saturation is KHA = 2.7 x 10(-4) M. Using these values the membrane conductance induced by the ion carrier nonactin and the shape of the current-voltage relationship as a function of the ligand concentration in water was analyzed. A maximum dipole potential of VDmax = -239 mV and a contribution of b = 3.1 x 10(-15) V cm2 per single adsorbed 2,4-D molecule was found. 74% of the dipole potential acts on the inner membrane barrier separating the two interfacial adsorption planes of nonactin. The remainder (26%) favours interfacial complex formation between nonactin and K+ from the aqueous phase. The data hold for membranes formed from dioleoyllecithin in n-decane.

A laser-temperature-jump method for the study of the rate of transfer of hydrophobic ions and carriers across the interface of thin lipid membranes

The first application of a laser-temperature-jump apparatus for the study of ion transport through planar (artificial) lipid membranes is described. The relaxation of the electric current is detected, either continuously at a constant applied voltage or discontinuously by a series of short voltage pulses. The second technique, a combined voltage- and temperature-jump method, is especially appropriate to investigate the kinetics of the adsorption/desorption process of hydrophobic ions and neutral carriers of cations at the membrane interface and to separate this phenomenon from the diffusion process through the unstirred aqueous layers adjacent to the membrane. The aim is to determine the rate-limiting step of transport. The permeation rate of the hydrophobic anion 2,4,6-trinitrophenolate is limited by the inner membrane barrier. For tetraphenylberate the rate constant of translocation across the inner barrier and that of desorption from the membrane into water are found to be of comparable magnitude. The membrane permeability of the neutral macrocyclic ion carrier enniatin B is strongly interface limited by its comparatively small rate of desorption into water. These results show that the frequently used a priori assumption of partition equilibrium at the membrane interfaces during transport is not justified.

Phase Transition in Di-oleoylphosphatidylglycerol/Monoolein Membranes due to Interactions of Positively Charged Peptides at their Lipid Membrane-Interface

To elucidate the factors that induce phase transitions in biomembranes due to interactions of proteins/peptides at the lipid membrane-interface, the effects of positively charged peptides on the cubic phase (Q229) of Dioleoylphosphatidylglycerol (DOPG)/Monoolein (MO) membranes were investigated. Small angle X-ray Scattering (SAXS) results revealed that 12 mol% DOPG/88 mol % MO membranes in excess water at 25°C is body centered cubic phase of crystallographic space group Im3m (Q229). In presence of peptide LLKKK, the lattices constant of Q229 phase was gradually decreased with an increase of peptide concentration and a phase transition from cubic (Q229) to cubic (Q224) phase occurred at R=0.080; (R= molar ratio of peptide to lipid). On the other hand the designed peptide WLFLLKKK and antimicrobial peptide Magainin-2 induced lamellar phase (Lα) in the same mixture membranes. These results indicate that the interactions of the these peptides with this mixture membrane are different: LLKKK induces electrostatic attractive interactions and that of WLFLLKKKK and Magainin-2 bound with the lipid membranes induce electrostatic repulsive interaction at the membrane-interface, might be the major factor inducing different phase transitions in 12 mol% DOPG/88mol% MO mixture membranes. Key words: Antimicrobial peptide Magain-2; Dioleoylphosphatidylglycerol; Monoolein; Cubic phases; Small angle X-ray Scattering DOI: 10.3329/bjsir.v45i3.6530Bangladesh J. Sci. Ind. Res. 45(3), 219-224, 2010