Lipid rafts of mouse liver contain nonextended and extended acetylcholinesterase variants along with M3 muscarinic receptors

The observation of acetylcholinesterase (AChE) type H (AChEH), which is the predominant AChE variant in visceral organs and immune cells, in lipid rafts of muscle supports functional reasons for the raft targeting of glypiated AChEH. The search for these reasons revealed that liver AChE activity is mostly confined to rafts and that the liver is able to make N‐extended AChE variants and target them to rafts. These results prompted us to test whether AChE and muscarinic receptors existed in the same raft. Isolation of flotillin2‐rich raft fractions by their buoyancy in sucrose gradients, followed by immunoadsorption and matrix‐assisted laser desorption ionization‐time of flight‐mass spectrometry application, gave the following results: 1) most hepatic AChE activity emanates from AChE‐H mRNA, and its product, glypiated AChEH, accumulates in rafts; 2) N‐extended N‐AChE readthrough variant, nonglypiated N‐AChEH, and N‐AChE tailed variant were all identified in liver rafts; and 3) M3 AChRs were observed in rafts, and coprecipitation of raft‐confined N‐AChE and M3 receptors by using anti‐M3 antibodies showed that enzyme and receptor reside in the same raft unit. A raft domain that harbors tightly packed muscarinic receptor and AChE may represent a molecular device that, by means of which, the intensity and duration of cholinergic inputs are regulated.—Montenegro, M. F., Cabezas‐Herrera, J., Campoy, F. J., Muñoz‐Delgado, E., Vidal, C. J. Lipid rafts of mouse liver contain non‐extended and extended acetylcholinesterase variants along with M3 muscarinic receptors. FASEB J. 31, 544–555(2017). www.fasebj.org