Abstract
The phosphorylation of connexins represents an important mechanism that regulates the biological activity of gap junctions. The methods described in this chapter to study connexin phosphorylation utilize [32P]orthophosphate metabolic radiolabeling of intact cells which permits the subsequent direct identification of phosphorylated amino acids and phosphopeptides of connexin. These methods include: (1) the analysis of connexin phosphoisoforms by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), (2) direct identification of the phosphorylated amino acid(s) by two-dimensional phosphoamino acid analysis, and (3) resolution of phosphorylated connexin peptides by two-dimensional phosphotryptic peptide analysis. Connexin43 (Cx43) is used as the primary experimental example in this chapter because of the authors’ extensive experience with this connexin subtype.
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© 2001 Humana Press Inc.
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Warn-Cramer, B.J., Kurata, W.E., Lau, A.F. (2001). Biochemical Analysis of Connexin Phosphorylation. In: Bruzzone, R., Giaume, C. (eds) Connexin Methods and Protocols. Methods In Molecular Biology™, vol 154. Humana Press. https://doi.org/10.1385/1-59259-043-8:431
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DOI: https://doi.org/10.1385/1-59259-043-8:431
Publisher Name: Humana Press
Print ISBN: 978-0-89603-658-1
Online ISBN: 978-1-59259-043-8
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