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Related Concept Videos

MAPK Signaling Cascades01:07

MAPK Signaling Cascades

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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When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze the...
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Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as  SH2...
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Microtubule function and architecture are regulated by an array of specialized proteins called microtubule-associated proteins or MAPs. These proteins are widespread across different organisms and have conserved protein motifs, like the multi-TOG domain for tubulin binding found in the CLASP family of MAPs. Some MAPs are lineage-specific based on their conserved domains. Their functions depend upon the cytoskeletal architecture and cell type they are located within. In-plant cells, a specific...
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Assaying Protein Kinase Activity with Radiolabeled ATP
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Published on: May 26, 2017

Structural studies of MAP Kinase cascade components.

Elizabeth J Goldsmith1, Xiaoshan Min, Haixia He

  • 1Department of Biochemistry, The University of Texas Southwestern Medical Center at Dallas, Dallas, TX, USA. Elizabeth.Goldsmith@UTSouthwestern.edu

Methods in Molecular Biology (Clifton, N.J.)
|September 3, 2010
PubMed
Summary
This summary is machine-generated.

Structural analysis of MAPK cascade components revealed unique inactive kinase conformers. Selenomethionine substitution in MAP2K MEK6 and MAP3K TAO2 was used for de novo phasing, aiding crystal structure determination.

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Published on: July 17, 2019

Area of Science:

  • Structural biology
  • Molecular enzymology
  • Signal transduction pathways

Background:

  • Mitogen-activated protein kinase (MAPK) cascades are crucial for cellular signaling.
  • Understanding MAPK enzyme activation and interaction is vital.
  • Many MAPK kinases adopt unique inactive conformations, complicating structural studies.

Purpose of the Study:

  • To investigate the structural properties of MAPK cascade components.
  • To facilitate de novo phasing for crystal structure determination of specific kinases.
  • To explore the utility of selenomethionine (SeMet) substitution in structural studies of MAP2K MEK6 and MAP3K TAO2.

Main Methods:

  • Preparation and expression of MAP2K MEK6 and MAP3K TAO2.
  • Incorporation of selenomethionine (SeMet) into TAO2 for experimental phasing.
  • Expression of wild-type TAO2 and SeMet-substituted TAO2 in insect cells.

Main Results:

  • Successful expression of both wild-type TAO2 and SeMet-substituted TAO2.
  • Demonstrated feasibility of using SeMet substitution for de novo phasing of these kinases.
  • Provided a foundation for future high-resolution structural determination.

Conclusions:

  • Selenomethionine substitution is an effective strategy for solving the crystal structures of MAPK kinases like MEK6 and TAO2.
  • This approach overcomes limitations of molecular replacement for kinases adopting unique inactive conformers.
  • The study advances structural insights into MAPK signaling regulation.