Analysis of mitogen-activated protein kinase activation and interactions with regulators and substrates
Open accessAbstract
Mitogen-activated protein kinase (MAPK) cascades are ubiquitous signal transduction modules in eukaryotes that are of great interest and importance. Here, we summarize some useful methods for the analysis of MAPK signaling, including methods to (1) detect MAPK activation in cells, with an emphasis on using phosphorylation-state-specific antibodies raised against mammalian phosphopeptide sequences to detect the activation of MAPKs in other species; (2) estimate the cellular concentrations of MAPKs and other proteins of interest; (3) detect and quantify the stable physical association of MAPKs with their substrates and regulators, and estimate the relevant dissociation constants; (4) delineate the MAPK-binding regions or domains of MAPK-interacting proteins, with particular emphasis on the identification and verification of MAPK-docking sites. These procedures are broadly applicable to many organisms, including both yeast and mammalian cells.
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The structural and biochemical analysis, together with other studies, clarified the minimal sequence requirements for linear motifs to bind the MAPK docking groove.
“Our structural and biochemical analysis in combination with other studies clarified the minimal sequence requirements for linear motifs to bind the MAPK docking groove (42-47).”
From Specificity of Linear Motifs That Bind to a Common Mitogen-Activated Protein Kinase Docking Groove (2012) · cites this paper as [42]
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Specificity of Linear Motifs That Bind to a Common Mitogen-Activated Protein Kinase Docking Groove
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- Authors
- BARDWELL, L., & SHAH, K.
- Affiliations
- University of California, Irvine (US)
- Corresponding author
- BARDWELL, L
- ORCID
- BARDWELL, L
- Venue
- Methods
- Publisher
- Elsevier BV
- Published
- 2006-11
- Volume
- 40
- Issue
- 3
- Pages
- 213-223
- ISSN
- 1046-2023 (ISSN-L)
- PMC
- PMC3017500
- OpenAlex
- W1970071313
- Cited by
- 46
- References
- 80
- Citations by year
- 2026: 4 · 2025: 1 · 2023: 2 · 2022: 3 · 2021: 5 · 2020: 4 · 2018: 3 · 2017: 1 · 2015: 1 · 2014: 4 · 2013: 3 · 2012: 3
- FWCI
- 0.54
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Methods (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of General Medical Sciences (GM-60366, R33GM069013, R01GM060366, GM69013, R21GM069013)
- 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 › Protein Tyrosine Phosphatases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Melanoma and MAPK Pathways
- Keywords
- mitogen-activated protein kinase, MAPK signaling, MAPK substrates, protein-protein interaction, dissociation constant, signal transduction
- MeSH terms
- Amino Acid Sequence, Animals, Antibody Specificity, Binding Sites, Buffers, Cell Extracts, Enzyme Activation, Humans, Molecular Biology, Molecular Sequence Data, Phosphorylation, Proteins, Mitogen-Activated Protein Kinases, Immunoprecipitation
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