Aqueous Polymer Two-phase Partition for The Proteomic Analysis of Plasma Membrane From Rat Dorsal Root Ganglion Neurons*

Abstract

Dorsal root ganglion (DRG) neurons are primary sensory neurons that conduct neuronal impulses related to pain,touch and temperature senses.To comprehensively identify proteins of plasma membrane (PM) from small amount of dorsal root ganglion (DRG) neurons,a proteomics strategy that utilizes aqueous polymer two-phase partition in combination with differential velocity centrifugation was adopted to enrich the PM,followed by SDS-PAGE,CapLC-MS/MS and bioinformatics analysis.Western blot analysis showed that the concentration of PM in purified plasma membrane(PPM) was 2.3 times higher than that in crude plasma membrane(CPM),15 times higher than that in whole tissue lysate (WTL).By searching against the rat IPI protein sequence database,a total of 729 non-redundant proteins were identified from the PM preparation,of which 547 had a gene ontology (GO) annotation indicating a cellular component,and 159 (21.8%) were unambiguously identified as PM proteins.A data set of plasma membrane proteins of DRG as well as a tool to study PM proteins were provided in a small amounts of sample.