title: Nonideal mixing in multicomponent lipid/detergent systems.
abstract: A detailed understanding of the mixing properties of membranes to which detergents are added is mandatory for improving the application and interpretation of detergent based protein or lipid extraction assays. For Triton X-100 (TX-100), a nonionic detergent frequently used in the process of solubilizing and purifying membrane proteins and lipids, we present here a detailed study of the mixing properties of binary and ternary lipid mixtures by means of high-sensitivity isothermal titration calorimetry (ITC). To this end the partitioning thermodynamics of TX-100 molecules from the aqueous phase to lipid bilayers composed of various mixtures of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), egg-sphingomyelin (SM), and cholesterol (cho) are characterized. Composition-dependent partition coefficients K are analysed within the frame of a thermodynamic model developed to describe nonideal mixing in multicomponent lipid/detergent systems. The results imply that POPC, fluid SM, and TX-100 mix almost ideally (nonideality parameters |ρ(α/β)|<RT). However, favourable SM/cho (ρ(SM/cho)≤-6RT) and unfavourable PC/cho interactions (ρ(PC/cho) = 2RT) may under certain conditions cause POPC/TX-100-enriched domains to segregate from SM/cho-enriched ones. TX-100/cho contacts are unfavourable (ρ(cho/TX) = 4RT), so the system tends to avoid them. That means, addition of TX-100 promotes the separation of SM/cho-rich from PC/TX-100-rich domains. It appears that cho/detergent interactions are crucial governing the abundance and composition of detergent-resistant membrane patches.
authors.affiliation: Biozentrum of the University of Basel, Division of Biophysical Chemistry, Klingelbergstrasse 70, CH-4056 Basel, Switzerland.
authors.forename: Alekos
authors.initials: A
authors.lastname: Tsamaloukas
authors.forename: Halina
authors.initials: H
authors.lastname: Szadkowska
authors.forename: Heiko
authors.initials: H
authors.lastname: Heerklotz