Identification of ciliary neurotrophic factor receptor alpha as a mediator of neurotoxicity induced by extracellular alpha-synuclein

MES cells were treated with vehicle or 250 nM extracellular alpha-synuclein and plasma membrane proteins were labeled with biotin, followed by affinity purification. The biotinylated enriched membranous proteins were incubated with an alpha-synuclein peptide array membrane, washed extensively with TBS-T, followed by incubation with avidin-HRP. The positive spots were excised and the bound proteins were eluted. The elutions from consecutive spots were combined before trypsin digestion and analysis of peptides by LC-MS/MS.