A myristoylated peptide corresponding to the N-terminus of NAP-22 (neuronal axonal myristoylated membrane protein of 22 kDa) causes the quenching of the fluorescence of BODIPY®-TMR-labelled PtdIns(4,5)P2 in bilayers of 1-palmitoyl-2-oleoyl phosphatidylcholine containing 40 mol% cholesterol and 0.1 mol% BODIPY®–PtdIns(4,5)2. Both fluorescence spectroscopy and total internal reflectance fluorescence microscopy revealed the cholesterol-dependent nature of PtdIns(4,5)P2-enriched membrane-domain formation.

Abbreviations used: BODIPY®-FL, 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl-aminohexanoyl; BODIPY®-TMR, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene-1,3-dimethyl-5-(4-methoxyphenyl)-2-propionyl-aminohexanoyl-aminohexanoyl; CAP-23, cortical cytoskeleton-associated protein of 23 kDa; GAP-43, growth-associated protein of 43 kDa; LUV, large unilamellar vesicle; MARCKS, myristoylated alanine-rich C-kinase substrate; NAP-22, neuronal axonal myristoylated membrane protein of 22 kDa; POPC, 1-palmitoyl-2-oleoyl phosphatidylcholine; POPE, 1-palmitoyl-2-oleoyl phosphatidylethanolamine; PLC, phospholipase C; TIRF, total internal reflectance fluorescence.

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