Connexin multi-site phosphorylation: mass spectrometry-based proteomics fills the gap.

Connexins require an integrated network for protein synthesis, assembly, gating, internalization, degradation and feedback control that are necessary to regulate the biosynthesis, and turnover of gap junction channels. At the most fundamental level, the introduction of sequence-altering, modifications introduces changes in protein conformation, activity, charge, stability and localization. Understanding the sites, patterns and magnitude of protein post-translational modification, including phosphorylation, is absolutely critical. Historically, the examination of connexin phosphorylation has been placed within the context that one or small number of sites of modification strictly corresponds to one molecular function. However, the release of high-profile proteomic datasets appears to challenge this dogma by demonstrating connexins undergo multiple levels of multi-site phosphorylation. With the growing prominence of mass spectrometry in biology and medicine, we are now getting a glimpse of the richness of connexin phosphate signals. Having implications to health and disease, this review provides an overview of technologies in the context of targeted and discovery proteomics, and further discusses how these techniques are being applied to “fill the gaps” in understanding of connexin post-translational control. This article is part of a Special Issue entitled: The Communicating junctions, roles and dysfunctions.