title: Label-free differential analysis of murine postsynaptic densities.
abstract: This chapter provides detailed methodology for the enrichment and label-free differential analysis of postsynaptic density (PSD) proteins. Methods discussed will include tissue homogenization, subcellular fractionation, protein digestion, and label-free differential analysis after liquid chromatography-tandem mass spectrometry. When combined, these protocols facilitate the identification of receptors and signal transducers that comprise the PSD and provide an optimized workflow for the differential analysis of PSD proteomes. This strategy supports a utility for coupling fractionation with proteomics analysis to enrich for low-abundant proteins in cellular localizations that would otherwise be lost in a global tissue context.
authors.affiliation: University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
authors.forename: Scott P
authors.initials: SP
authors.lastname: Goulding
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Maccoss
authors.forename: Christine C
authors.initials: CC
authors.lastname: Wu
mesh_terms: D000818:Animals; D002499:Centrifugation, Density Gradient; D002853:Chromatography, Liquid; D051379:Mice; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000737:chemistry; D057907:Post-Synaptic Density / Q000737:chemistry*; D020543:Proteome / Q000032:analysis; D040901:Proteomics / Q000379:methods*; D013347:Subcellular Fractions / Q000737:chemistry; D053719:Tandem Mass Spectrometry
chemical_list: D009419:Nerve Tissue Proteins; D020543:Proteome; C033652:postsynaptic density proteins