Mass spectrometry-based phosphoproteomics reveals multisite phosphorylation on mammalian brain voltage-gated sodium and potassium channels

Abstract

Voltage-gated sodium and potassium channels underlie electrical activity of neurons, and are dynamically regulated by diverse cell signaling pathways that ultimately exert their effects by altering the phosphorylation state of channel subunits. Recent mass spectrometric-based studies have led to a new appreciation of the extent and nature of phosphorylation of these ion channels in mammalian brain. This has allowed for new insights into how neurons dynamically regulate the localization, activity and expression through multisite ion channel phosphorylation.

Figure 1

Flowchart for identification/characterization of phosphorylation in VGSCs and VGKCs. Antibody-dependent (A, B) and -independent (C) approaches are shown (see text for details). A. Conventional antibody-dependent method for purification of target proteins from a complex sample. B. Enrichment of phosphopeptides from a complex sample using a phosphospecific antibody (Filter aided antibody capturing and elution: FACE). C. High-throughput strategy for phosphorylation identification from complex samples in the absence of an antibody (FASP: filter aided sample preparation)