title: Interaction of saponin 1688 with phase separated lipid bilayers.
abstract: Saponins are a diverse family of naturally occurring plant triterpene or steroid glycosides that have a wide range of biological activities. They have been shown to permeabilize membranes and in some cases membrane disruption has been hypothesized to involve saponin/cholesterol complexes. We have examined the interaction of steroidal saponin 1688-1 with lipid membranes that contain cholesterol and have a mixture of liquid-ordered (Lo) and liquid-disordered (Ld) phases as a model for lipid rafts in cellular membranes. A combination of atomic force microscopy (AFM) and fluorescence was used to probe the effect of saponin on the bilayer. The results demonstrate that saponin forms defects in the membrane and also leads to formation of small aggregates on the membrane surface. Although most of the membrane damage occurs in the liquid-disordered phase, fluorescence results demonstrate that saponin localizes in both ordered and disordered membrane phases, with a modest preference for the disordered regions. Similar effects are observed for both direct incorporation of saponin in the lipid mixture used to make vesicles/bilayers and for incubation of saponin with preformed bilayers. The results suggest that the initial sites of interaction are at the interface between the domains and surrounding disordered phase. The preference for saponin localization in the disordered phase may reflect the ease of penetration of saponin into a less ordered membrane, rather than the actual cholesterol concentration in the membrane. Dye leakage assays indicate that a high concentration of saponin is required for membrane permeabilization consistent with the supported lipid bilayer experiments.
authors.affiliation: Measurement Science and Standards, National Research Council Canada, Ottawa, ON K1A 0R6, Canada.
authors.forename: Maohui
authors.initials: M
authors.lastname: Chen
authors.affiliation: Measurement Science and Standards, National Research Council Canada, Ottawa, ON K1A 0R6, Canada.
authors.forename: Vinod
authors.initials: V
authors.lastname: Balhara
authors.affiliation: Measurement Science and Standards, National Research Council Canada, Ottawa, ON K1A 0R6, Canada.
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Jaimes Castillo
authors.affiliation: Aquatic and Crop Resource Development, National Research Council Canada, Saskatoon, SK S7N 0W9, Canada.
authors.forename: John
authors.initials: J
authors.lastname: Balsevich
authors.affiliation: Measurement Science and Standards, National Research Council Canada, Ottawa, ON K1A 0R6, Canada. Electronic address: Linda.Johnston@nrc-cnrc.gc.ca.
authors.forename: Linda J
authors.initials: LJ
authors.lastname: Johnston
mesh_terms: D002463:Cell Membrane Permeability; D008051:Lipid Bilayers / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D012503:Saponins / Q000737:chemistry*
chemical_list: D008051:Lipid Bilayers; D012503:Saponins
keywords: Atomic force microscopy; Dye leakage; Fluorescence; Lipid bilayer; Saponin