Search

Membrane organization and fluidity research continues to expand and the understanding of membrane dynamics continues to be refined. Within this field of study, laurdan remains among the most popular, versatile, and established fluorescence probes. Fluorimetry and multiphoton microscopy techniques are standards for measuring laurdan fluorescence and continue to be refined. However, complications have arisen due to an amended membrane model, revised terms used for describing membrane phases, and wide variation in the selection of laurdan generalized polarization equation values. Here, in the context of the history and chemical properties of laurdan, discrepancies are highlighted and important recommendations are made to promote uniformity and ongoing progress. [ABSTRACT FROM AUTHOR]

Cholesterol-enriched functional portions of plasma membranes, such as caveolae and rafts, were isolated from lungs of wild-type (WT) and caveolin-1 knockout (Cav-1 KO) mice within detergent resistant membranes (DRMs). To gain insight into their molecular composition we performed proteomic and lipid analysis on WT and Cav-1 KO-DRMs that showed predicted variations of proteomic profiles and negligible differences in lipid composition, while Langmuir monolayer technique and small and wide-angle X-ray scattering (SAXS-WAXS) were here originally introduced to study DRMs biophysical association state. Langmuir analysis of Cav-1 containing DRMs displayed an isotherm with a clear-cut feature, suggesting the coexistence of the liquid-ordered (Lo) phase typical of the raft structure, namely "cholesterol-rich Lo phase", with a phase fully missing in Cav-1 KO that we named "caveolin-induced Lo phase". Furthermore, while the sole lipid component of both WT and KO-DRMs showed qualitatively similar isotherm configuration, the reinsertion of recombinant Cav-1 into WT-DRMs lipids restored the WT-DRM pattern. X-ray diffraction results confirmed that Cav-1 causes the formation of a "caveolin-induced Lo phase", as suggested by Langmuir experiments, allowing us to speculate about a possible structural model. These results show that the unique molecular link between Cav-1 and cholesterol can spur functional order in a lipid bilayer strictly derived from biological sources. [ABSTRACT FROM AUTHOR]