Calcium triggered Lα-H2 phase transition monitored by combined rapid mixing and time-resolved synchrotron SAXS

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Abstract

Background: Awad et al. [1] reported on the Ca2+-induced transitions of dioleoyl-phosphatidylglycerol (DOPG)/monoolein (MO) vesicles to bicontinuous cubic phases at equilibrium conditions. In the present study, the combination of rapid mixing and time-resolved synchrotron small angle X-ray scuttering (SAXS) was applied for the in situ investigations of fast structural transitions of diluted DOPG/MO vesicles into well-ordered nanostructures by the addition of low concentration Ca2+ solutions. Methodology/Principal Findings: Under static conditions and the in absence of the divalent cations, the DOPG/MO system forms large vesicles composed of weakly correlated bilayers with a d-spacing of ∼ 140 Å (Lα-phase). The utilization of a stopped-flow apparatus allowed mixing these DOPG/MO vesicles with a solution of Ca2+ ions within 10 milliseconds (ms). In such a way the dynamics of negatively charged PG to divalent cation interactions, and the kinetics of the induced structured transitions were studied. Ca2+ ions have a very strong impact on the lipidic nanostructure. Intriguingly, already at low salt concentrations (DOPG/Ca2+ > 21, Ca2+ ions trigger the transformation from bilayers to monolayer nanotubes unverted hexagonal phase, H2). Our results reveal that a binding ratio of 1 Ca2+ per 8 DOPG is sufficient for the formation of the H2 phase. At 50°C a direct transition from the vesicles to the H2 phase was observed, whereas at ambient temperature (20°C) a short lived intermediate phase (possibly the cubic Pn3m phase) coexisting with the H2 phase was detected. Conclusions/Significance: The strong binding of the divalent cations to the negatively charged DOPG molecules enhances the negative spontaneous curvature of the monolayers and causes a rapid cullapsing of the vesicles. The rapid loss of the bilayer stability and the reorganization of the lipid molecules within ms support the argument that the transition mechanism is based on a leoky fusion of the vesicles. © 2008 Yaghmur et al.

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Yaghmur, A., Laggner, P., Sartori, B., & Rappolt, M. (2008). Calcium triggered Lα-H2 phase transition monitored by combined rapid mixing and time-resolved synchrotron SAXS. PLoS ONE, 3(4). https://doi.org/10.1371/journal.pone.0002072

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