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Related Concept Videos

MAPK Signaling Cascades01:07

MAPK Signaling Cascades

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Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
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Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays
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Analysis of MAPK activities using MAPK-specific antibodies.

Roland Willmann1, Daniel J Haischer, Andrea A Gust

  • 1Department of Plant Biochemistry, ZMBP, University of Tübingen, Auf der Morgenstelle 32, 72076, Tübingen, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|June 9, 2014
PubMed
Summary
This summary is machine-generated.

Mitogen-activated protein (MAP) kinase assays are crucial for understanding cellular stress responses. New methods are needed for unambiguous identification of activated MAP kinases, as current techniques lack specificity.

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Area of Science:

  • Cellular biology
  • Molecular biology
  • Biochemistry

Background:

  • Mitogen-activated protein kinases (MAPKs) regulate critical cellular processes like stress response through protein phosphorylation.
  • Accurate assays for MAPK activity are essential research tools.

Purpose of the Study:

  • To address the limitations of current methods in unambiguously identifying activated MAP kinases.
  • To highlight the need for specific purification techniques.

Main Methods:

  • Review of existing techniques including in-gel kinase assays and phospho-specific western blots.
  • Discussion of immunoprecipitation for kinase purification.

Main Results:

  • In-gel kinase assays and phospho-specific western blots do not provide unambiguous identification of activated MAPKs.
  • Immunoprecipitation allows for specific purification of the target kinase.

Conclusions:

  • Current MAPK assays lack specificity for identifying individual activated kinases.
  • Immunoprecipitation prior to analysis is recommended for unambiguous MAPK identification.