Proteomic investigation of human α7-nicotinic acetylcholine receptor interacting proteins (1147.5)

The α7-nicotinic acetylcholine receptor (α7-nAChR) is a ligand-gated ion channel widely expressed in the mammalian CNS that is associated with numerous physiological functions. α7-nAChRs are also the principal high-affinity α-bungarotoxin (bgtx) binding proteins in the mammalian brain. There is evidence that α7-nAChRs are involved in a number of protein-protein signaling cascades. SH-SY5Y and SH-EP1 human clonal cell lines have been utilized extensively to characterize nAChRs. SH-SY5Y cells naturally express α7-nAChRs. SH-EP1-hα7 and SH-EP1-hα7-Ric-3 cells have been engineered to stably and heterologously express human α7-nAChRs alone or along with human Resistance to Inhibitors of Cholinesterase 3 (Ric-3). Bgtx-sensitive protein complexes were isolated from each cell line using bgtx affinity immobilization. Bound protein was eluted, reduced and alkylated prior to digestion with trypsin in-solution. The resulting peptides were analyzed with an LTQ-Orbitrap mass spectrometer and data analyzed using ProteoI...