Quantitative Analysis of Plasma Membrane Proteome Using Two-Dimensional Difference Gel Electrophoresis

The plasma membrane (PM) controls cell’s exchange of both material and information with the outside environment, and PM-associated proteins play key roles in cellular regulation. Numerous cell surface receptors allow cells to perceive and respond to various signals from neighbor cells, pathogens, or the environment; large numbers of transporter and channel proteins control material uptake or release. Quantitative proteomic analysis of PM-associated proteins can identify key proteins involved in signal transduction and cellular regulation. Here, we describe a protocol for quantitative proteomic analysis of PM proteins using two-dimensional difference gel electrophoresis. The protocol has been successfully employed to identify new components of the brassinosteroid signaling pathway, and should also be applicable to the studies of other plant signal transduction pathways and regulatory mechanisms.