Docking Motif Interactions in MAP Kinases Revealed by Hydrogen Exchange Mass Spectrometry
Open accessAbstract
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 beta7-beta8 and alphaD-alphaE in p38alpha 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, alphaF 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.
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Earlier H/D-MS studies from the Ahn laboratory show that the dynamics of key loops are affected by the interactions of MAPKs with peptides and ligands.
“This supports the results of earlier H/D-MS studies from the Ahn laboratory that show the dynamics of key loops are affected be the interactions of MAPKs with peptides and ligands.66”
From Molecular basis of MAP kinase regulation (2013) · cites this paper as [66]
Allostery among the catalytic site, activation loop, and peptide-docking site is well documented for ERK and p38 MAPKs but is less understood for JNK.
“Allostery among the catalytic site, A-loop, and peptide-docking site of ERK and p38 MAPKs has been well documented (Chang et al., 2002; Goldsmith, 2011; Lee et al., 2004; Wang et al., 1997; Zhang et al., 1994; Zhou et al., 2006a, 2006b), but much less is known about these processes for JNK.”
From Structural Mechanisms of Allostery and Autoinhibition in JNK Family Kinases (2012) · cites this paper as Lee et al., 2004
FxFP motifs in certain MAPK substrates and a separate docking groove for such motifs on ERK suggest that two docking grooves on ERK may operate in combination; Elk1 contains both an FxFP and a D-motif.
“Moreover, the existence of FxFP (Phe-any amino acid-Phe-Pro) motifs in certain MAPK (especially ERK) substrates and a separate docking groove for such motifs on ERK suggest that in some cases two docking grooves on ERK may operate in combination: Elk1 for example contains both an FxFP and a D-motif (35-37).”
From Specificity of Linear Motifs That Bind to a Common Mitogen-Activated Protein Kinase Docking Groove (2012) · cites this paper as [35]
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- Authors
- Lee, T., Hoofnagle, A. N., Kabuyama, Y., Stroud, J., Min, X., Goldsmith, E. J., Chen, L., Resing, K. A., & Ahn, N. G.
- Affiliations
- University of Colorado Boulder (US)
- Howard Hughes Medical Institute (US)
- The University of Texas Southwestern Medical Center (US)
- Corresponding author
- Ahn, Natalie G
- ORCID
- Venue
- Molecular Cell
- Publisher
- Elsevier BV
- Published
- 2004-04
- Volume
- 14
- Issue
- 1
- Pages
- 43-55
- ISSN
- 1097-2765 (ISSN-L)
- OpenAlex
- W2020513008
- Cited by
- 282
- References
- 35
- Citations by year
- 2026: 3 · 2025: 6 · 2024: 5 · 2023: 12 · 2022: 8 · 2021: 8 · 2020: 11 · 2019: 9 · 2018: 10 · 2017: 11 · 2016: 17 · 2015: 14 · 2014: 9 · 2013: 9 · 2012: 15
- FWCI
- 9.02
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Molecular Cell (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of General Medical Sciences (R01GM064642, R01-GM48521, R01GM048521)
- National Institute of Allergy and Infectious Diseases (R21AI049905)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Melanoma and MAPK Pathways
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Keywords
- mitogen-activated protein kinase, p38α, ERK2, protein-protein interaction, hydrogen-deuterium exchange mass spectrometry, substrate binding, Elk-1, nucleoporins, activation loop
- MeSH terms
- Amino Acid Sequence, Animals, Binding Sites, Enzyme Activation, Models, Molecular, Molecular Sequence Data, Mutation, Peptides, Protein Binding, Mass Spectrometry, Sequence Alignment, Protein Structure, Tertiary, Mitogen-Activated Protein Kinases, Rats
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