Simplified enrichment of plasma membrane proteins for proteomic analyses in Arabidopsis thaliana

Proteomics. 2011 May;11(9):1780-8. doi: 10.1002/pmic.201000648. Epub 2011 Mar 23.

Abstract

The plasma membrane (PM) serves as the point of contact between cells and the outside environment. As such, changes in the PM proteome are an important component of understanding cellular responses to a diverse array of stimuli. However, intricate sample handling to enrich PM proteomes by traditional methods is both technically challenging and time consuming. Here, we describe a simplified method for decreasing the representation of other membrane-containing organelles such as the endoplasmic reticulum, plastids and mitochondria from crude microsomal membrane isolations. The decrease in other organellar proteomes results in an increase in both the total number of PM proteins and the number of spectra identified from these proteins representing the PM proteome. Therefore, this strategy represents a simple and rapid method for enriching PM proteins from Arabidopsis cell cultures for proteomic analyses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / cytology
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / analysis*
  • Arabidopsis Proteins / isolation & purification
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • Chromatography, Liquid
  • Electrophoresis, Polyacrylamide Gel / methods
  • Mass Spectrometry
  • Membrane Proteins / analysis*
  • Membrane Proteins / isolation & purification
  • Proteome / analysis*
  • Proteome / isolation & purification
  • Proteomics / methods

Substances

  • Arabidopsis Proteins
  • Membrane Proteins
  • Proteome