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Cover Picture: On-Bead Chemical Synthesis and Display of Phosphopeptides for Affinity Pull-Down Proteomics (ChemBioChem 4/2006)
The cover picture shows how a polymer bead travels through chemistry space from chemical synthesis of its phosphopeptide ligands to use in the affinity pull-down of 14-3-3 proteins. First a phosphopeptide is synthesized on a PEGA support, next the phosphopeptide ligands selectively bind 14-3-3 proteins from cell lysates, finally the proteins are released for proteomic analysis and identification by mass spectrometry. Peptide sequences containing two phosphorylation motifs of the proapoptotic protein Bad were used as baits in affinity pull-down experiments to determine their ability to bind intracellular 14-3-3 proteins. The support-bound Bad phosphopeptides were able to pull-down 14-3-3 proteins from cell lysates, and all seven members of the 14-3-3 family were identified by mass spectrometry. More about this fast and efficient on-bead method for affinity pull-down proteomics is reported by K. J. Jensen et al. on p. 623 ff.