Protein and protein-lipid membranes from solubilized erythrocyte ghosts

Summary Erythrocyte ghosts were solubilized by addition of acid 2-chloroethanol to an aqueous membrane suspension. The proteins were separated from the lipids by chromatography on Sephadex LH-20 (Zahler and Wallach, 1967). Both the solution of separated proteins and of the total membrane in chloroethanol-water can be spread at a benzene-water interface. By lowering a thin teflon plate with a small hole through this interface, one can form protein or protein-lipid films over the hole. After the benzene has evaporated stable thin membranes are formed which contain only protein or protein together with lipid. The morphology and thickness of these membranes were investigated by different electron microscopic techniques.     

Effect of polar carotenoids on the oxygen diffusion-concentration product in lipid bilayers. An EPR spin label study.

The oxygen diffusion-concentration product was determined in phosphatidylcholine (PC) bilayers from oxygen broadening of the spin label EPR spectra. The use of fatty acid spin labels makes it possible to do structural and oximetric measurements with the same sample. We find that polar carotenoids, zeaxanthin and violaxanthin, increase ordering of hydrocarbon chains in saturated (dimyristoyl-PC) and unsaturated (egg yolk PC) membranes and also significantly decrease the oxygen diffusion-concentration product in the hydrocarbon region of these membranes. At 25 degrees C in the presence of 10 mol% of carotenoids, the product is about 30% smaller than in pure PC membranes. Intercalation of carotenoids decreases the oxygen diffusion-concentration product in the central part of the bilayer and has little effect on the product in the polar head group region. In contrast, cholesterol molecules significantly reduce the product on and near the membrane surface, and do not change it (saturated PC) or increase it (unsaturated PC) in the middle of the bilayer (Subczynski, W.K., Hyde, J.S. and Kusumi, A. (1989) Proc. Natl. Acad. Sci. USA 86, 4474-4478). The decrease of oxygen diffusion-concentration product may be a mechanism of carotenoid protective activity, which should be effective in plant and animal cells in the light as well as in the dark.