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 journalArticle

203 Citations (Scopus)

Abstract

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
Volume14
Issue number1
DOIs
StatePublished - Apr 9 2004

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Hydrogen
Mass Spectrometry
Phosphotransferases
Nuclear Pore Complex Proteins
Nuclear Pore
Porins
Nuclear Proteins
Lip
Proteins
Phosphorylation
Peptides
butyl phosphorotrithioate
In Vitro Techniques

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Docking motif interactions in Map kinases revealed by hydrogen exchange mass spectrometry. / Lee, Thomas; Hoofnagle, Andrew N.; Kabuyama, Yukihito; Stroud, James; Min, Xiaoshan; Goldsmith, Elizabeth J.; Chen, Lin; Resing, Katheryn A.; Ahn, Natalie G.

In: Molecular Cell, Vol. 14, No. 1, 09.04.2004, p. 43-55.

Research output: Contribution to journalArticle

Lee, T, Hoofnagle, AN, Kabuyama, Y, Stroud, J, Min, X, Goldsmith, EJ, Chen, L, Resing, KA & Ahn, NG 2004, 'Docking motif interactions in Map kinases revealed by hydrogen exchange mass spectrometry', Molecular Cell, vol. 14, no. 1, pp. 43-55. https://doi.org/10.1016/S1097-2765(04)00161-3
Lee, Thomas ; Hoofnagle, Andrew N. ; Kabuyama, Yukihito ; Stroud, James ; Min, Xiaoshan ; Goldsmith, Elizabeth J. ; Chen, Lin ; Resing, Katheryn A. ; Ahn, Natalie G. / Docking motif interactions in Map kinases revealed by hydrogen exchange mass spectrometry. In: Molecular Cell. 2004 ; Vol. 14, No. 1. pp. 43-55.
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