A mutagenesis-based screen to rapidly identify phosphorylation sites in mitogen-activated protein kinase substrates.

@article{PalmForster2012AMS,
  title={A mutagenesis-based screen to rapidly identify phosphorylation sites in mitogen-activated protein kinase substrates.},
  author={Mieder Anthony Thomas Palm-Forster and Lennart Eschen-Lippold and Justin Lee},
  journal={Analytical biochemistry},
  year={2012},
  volume={427 2},
  pages={
          127-9
        }
}
Identification and characterization of protein phosphorylation sites often requires mass spectrometric analysis, which is not trivial or accessible to many laboratories. Here, a targeted strategy to mutagenize putative phosphorylation sites within mitogen-activated protein kinase (MAPK) substrates is described. This employs a combination of standard type II with type IIs restriction enzymes to rapidly create individual or multiple phosphorylation site mutant versions of kinase substrates with… CONTINUE READING
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