Probing lipid interactions of plasma membrane proteins: a micropatterning approach
published: July 9, 2018, recorded: May 2018, views: 363
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It is well established that lipids and proteins are not just independent components of the plasma membrane of eukaryotic cells but that their arrangement, dynamics and function are interdependent.Besides direct lipid-protein interaction, transmembrane proteins are thought to bind a shell of annular lipids, which are more or less tightly associated with the proteins. Furthermore, highly ordered nanoscopic membrane domains have been proposed to act to compartmentalize proteins and their interactions, but have thus far not been directly observed. In this study, we use protein micropatterning combined with single-molecule tracking to directly measure lipid-protein interactions in the plasma membrane of living cells: different fluorescently labelled transmembrane proteins of interest (POIs) were captured and enriched within well-defined areas in the plasma membrane, leaving regions depleted of POI, which function as reference areas. From the distribution and diffusion behaviour of lipids and proteins with respect to the POI patterns, we were able to conclude on the local membrane environment of the POI. We found that a palmitoylated protein based on the transmembrane domain of hemagglutinin (HA-mGFP) influences its membrane environment well beyond the size of the transmembrane helix. The same effect was observed for a palmitoylation-deficient mutant allowing us to rule out formation of a more ordered membrane domain around HA-mGFP as the cause for this apparently increased protein size.
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