Docking motif interactions in Map kinases revealed by hydrogen exchange mass spectrometry

Thomas Lee, Andrew N. Hoofnagle, Yukihito Kabuyama, James Stroud, Xiaoshan Min, Elizabeth J. Goldsmith, Lin Chen, Katheryn A. Resing, Natalie G. Ahn

Research output: Contribution to journalArticlepeer-review

211 Scopus citations


Protein interactions between MAP kinases and substrates, activators, and scaffolding proteins are regulated by docking site motifs, one containing basic residues proximal to Leu-X-Leu (DEJL) and a second containing Phe-X-Phe (DEF). Hydrogen exchange mass spectrometry was used to identify regions in MAP kinases protected from solvent by docking motif interactions. Protection by DEJL peptide binding was observed in loops spanning β7-β8 and αD-αE in p38α and ERK2. In contrast, protection by DEF binding to ERK2 revealed a distinct hydrophobic pocket for Phe-X-Phe binding formed between the P+1 site, αF helix, and the MAP kinase insert. In inactive ERK2, this pocket is occluded by intramolecular interactions with residues in the activation lip. In vitro assays confirm the dependence of Elk1 and nucleoporin binding on ERK2 phosphorylation, and provide a structural basis for preferential involvement of active ERK in substrate binding and nuclear pore protein interactions.

Original languageEnglish (US)
Pages (from-to)43-55
Number of pages13
JournalMolecular cell
Issue number1
StatePublished - Apr 9 2004

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology


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