Effect of chlorophyll a on the phase behavior of hydrated monogalactosyldiacylglycerols.

We have studied the effect of chlorophyll a (chl a) on the X-ray diffraction patterns and the appearance of freeze-fracture electron micrographs of aqueous dispersions of monogalactosyldiacylglycerols (MGDG), the most abundant lipid in the thylakoid membrane. In MGDG systems containing 0-18 mol% of chl a, the diffraction patterns indicate the presence of a well-ordered reverse hexagonal phase. When 30 mol% of chl a was incorporated into the MGDG, the low-angle X-ray diffraction lines of the hexagonal lattice were slightly broadened and were accompanied by additional weak lines. With higher mol percents of chl a, the low-angle lines could no longer be indexed on a hexagonal or lamellar lattice. The freeze-fracture electron micrographs of similar samples showed that the patterns characteristic of the reverse hexagonal phase of an aqueous dispersion of pure MGDG were replaced by large liposomes, the fracture pattern of which is circular. We conclude that chl a in excess of 20 mol% destabilized the orderly reverse hexagonal phase of aqueous MGDG dispersions and disturbed the long-range order of the lipid array. These results are summarized in a temperature-composition isobaric phase diagram over a temperature range of -60 degrees C to 60 degrees C.

Statistical mechanics of lipid membranes. Protein correlation functions and lipid ordering.

An expression is derived for the lipid-mediated intermolecular interaction between protein molecules embedded in a lipid bilayer. It is assumed that protein particles are accommodated by the bilayer, but they distort the lipids in some manner from their equilibrium protein-free configuration. We treat this situation by expanding the free energy density in the plane of the membrane as a Taylor series in some arbitrary parameter and its gradient. Minimization of the total membrane energy for a given particle configuration yields the interparticle interaction energy for that configuration. A test of the model is provided by measurement of the protein-protein pair distribution function from freeze-fracture micrographs of partially aggregated membranes. The measured functions can be simulated by adjustment of two parameters (a) a lipid correlation length that characterizes the distance over which a distortion of the bilayers is transmitted laterally through the bilayer, and (b) a term quantifying the energy of the protein-lipid interaction at the protein-lipid boundary. Correlation lengths obtained by fitting the calculated particle distribution functions to the data are found to be several nanometers. Protein-lipid interaction energies are of the order of a few kT.

Pair distribution functions of bacteriorhodopsin and rhodopsin in model bilayers.

The pair distribution functions have been measured from freeze-fracture pictures of bacteriorhodopsin and rhodopsin recombinants with diacyl phosphatidylcholines (PC) of various hydrocarbon chain lengths. Pictures were used of samples that had been frozen from above the phase transition temperature of the lipid. Measured functions were compared with those calculated from two model interparticle potential energy functions, (a) a hard-disk repulsion only, and (b) a hard-disk repulsion plus electrostatic repulsion for a point charge buried in the membrane. The measured functions for bacteriorhodopsin di 12:0 PC, di 14:0 PC, and di 16:0 PC recombinants can be simulated using an interparticle hard-disk repulsion only. Bleached rhodopsin di 12:0 PC and di 18:1 trans-PC recombinants, and dark-adapted rhodopsin di 10:0 PC recombinants yield functions that are better simulated by assuming an additional repulsive interaction. The measured functions resemble those calculated using the hard-disk plus electrostatic repulsion model. The picture of dark-adapted rhodopsin in di 18:1 trans-PC frozen from 20 degrees C shows partial aggregation that is apparent in the measured pair distribution function. This attractive interaction persists even at 37 degrees C, where the measured function shows deviations from the hard-disk repulsive model, indicative of an attractive interparticle interaction. Implications of these results are discussed in terms of protein-lipid interactions.