Characterisation of plasmalemmal shedding of vesicles induced by the cholesterol/sphingomyelin binding protein, ostreolysin A-mCherry

摘要: Ostreolysin A (OlyA) is a 15-kDa protein that binds selectively to cholesterol/sphingomyelin membrane nanodomains. This binding induces the production of extracellular vesicles (EVs) comprise both microvesicles with diameters between 100nm and 1μm, larger around 10-μm diameter in Madin-Darby canine kidney cells. In this study, we show vesiculation these cells by fluorescent fusion OlyA-mCherry not affected temperature, mediated via intracellular Ca2+ signalling, does compromise cell viability ultrastructure. Seventy-one proteins are mostly cytosolic nuclear origin were detected shed EVs using mass spectroscopy. EVs, 218 84 lipid species identified, respectively, significantly enriched lysophosphatidylcholines cholesterol. Our collected data suggest nanodomains specific sorting into discrete patches, which promotes plasmalemmal blebbing EV shedding from We hypothesize effects accounted for changes local curvature upon OlyA-mCherry-plasmalemma interaction. potentially interesting model biophysical biochemical studies membranes, could represent tools non-invasive sampling thus metabolic fingerprinting.