The composition and nanoscale mechanical characteristics of the adhesive from two species of subaerial green unicellular microalgae (Chlorophyta), Coccomyxa sp. and Glaphyrella trebouxiodes, have been studied using Raman spectroscopy, chemical staining, and atomic force microscopy (AFM). Raman spectroscopy confirmed the adhesive proteins of both species to be predominantly in ß-sheet conformations and composed of a number of hydrophobic amino acid residues. Chemical staining with Congo red and thioflavin-T dyes further confirmed the presence of amyloid-like structures. Probing the adhesives with AFM revealed highly ordered and repetitive mechanical responses indicative of highly ordered structures within the adhesive. The repetitive nature of the sawtooth response is typical of a ''sacrificial bond'' and ''hidden length'' mechanism, and what wepropose is the result of mechanical manipulation of individual molecules within an intermolecular ß-sheet that makes up the generic amyloid structure. The mechanical data show how amyloid provides cohesive strength to the adhesives, and this intrinsic mechanical property of an amyloid-based adhesive explains the ecological success of attachment of these subaerial microalgae on various surfaces in urban environments. It is unknown to what extent amyloid fibrils occur in algal adhesives, but we postulate that the amyloid structure could provide a widespread mechanism for mechanical strength.
Tea tree oil (TTO) is the essential oil steam-distilled from Melaleuca alternifolia, a species of northern New South Wales, Australia. It exhibits a broad-spectrum antimicrobial activity and an antifungal activity. Only recently has TTO been shown to inhibit the in vitro growth of multidrug resistant (MDR) human melanoma cells. It has been suggested that the effect of TTO on tumor cells could be mediated by its interaction with the plasma membrane, most likely by inducing a reorganization of lipid architecture. In this paper we report biophysical and structural results obtained using simplified planar model membranes (Langmuir films) mimicking lipid "rafts". We also used flow cytometry analysis (FCA) and freeze-fracturing transmission electron microscopy to investigate the effects of TTO on actual MDR melanoma cell membranes. Thermodynamic (compression isotherms and adsorption kinetics) and structural (Brewster angle microscopy) investigation of the lipid monolayers clearly indicates that TTO interacts preferentially with the less ordered DPPC "sea" and that it does not alter the more ordered lipid "rafts". Structural observations, performed by freeze fracturing, confirm that TTO interacts with the MDR melanoma cell plasma membrane. Moreover, experiments performed by FCA demonstrate that TTO does not interfere with the function of the MDR drug transporter P-gp. We therefore propose that the effect exerted on MDR melanoma cells is mediated by the interaction with the fluid DPPC phase, rather than with the more organized "rafts" and that this interaction preferentially influences the ATP-independent antiapoptotic activity of P-gp likely localized outside "rafts".
Gangliosides are the main component of lipid rafts. These microdomains, floating in the outer leaflet of cellular membrane, play a key role in fundamental cellular functions. Little is still known about ganglioside and phospholipid interaction. We studied mixtures of dipalmitoylphosphatidylcholine and GD3 (molar fraction of 0.2, 0.4, 0.6, 0.8) using complementary techniques: 1), thermodynamic properties of the Langmuir-Blodgett films were assessed at the air-water interface (surface tension, surface potential); and 2), three-dimensional morphology of deposited films on mica substrates were imaged by atomic force microscopy. Mixture thermodynamics were consistent with data in the literature. In particular, excess free energy was negative at each molar fraction, thus ruling out GD3 segregation. Atomic force microscopy showed that the height of liquid-condensed domains in deposited films varied with GD3 molar fraction, as compatible with a lipid aggregation model proposed by Maggio. No distinct GD3-rich domain was observed inside the films, suggesting that GD3 molecules gradually mix with dipalmitoylphosphatidylcholine molecules, confirming DeltaG data. Morphological analysis revealed that the shape of liquid-condensed domains is strongly influenced by the amount of GD3, and an interesting stripe-formation phenomenon was observed. These data were combined with the thermodynamic results and interpreted in the light of McConnell's model.
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