Related Experiment Video
Updated: Aug 10, 2026
08:05
Assaying Protein Kinase Activity with Radiolabeled ATP
Published on: May 26, 2017
Analysis of mitogen-activated protein kinase activation
1Cardiovascular Research Institute, University of South Dakota School of Medicine, Sioux Falls, SD, USA.
Methods in Molecular Biology (Clifton, N.J.)
|August 20, 2005
Summary
Mast cell functions rely on protein kinases. New methods allow researchers to measure mitogen-activated protein kinase activation in single mast cells for better signal transduction studies.
Area of Science:
- Immunology and Cell Signaling
- Biochemistry and Molecular Biology
Background:
- Mast cells are crucial immune cells involved in allergic responses and inflammation.
- Their biological functions are tightly regulated by a complex network of protein kinases.
- Key kinases include tyrosine kinases (Fyn, Lyn, Syk, FAK) and serine/threonine kinases (Akt, PKC alpha/beta).
Purpose of the Study:
- To review recent advancements in quantifying mitogen-activated protein kinase (MAPK) activation.
- To highlight methodologies applicable to single mast cell analysis.
- To facilitate deeper understanding of signal transduction pathways in mast cells.
Main Methods:
- Focus on techniques for determining MAPK activation at the single-cell level.
- Discussion of methods applicable to studying extracellular signal-regulated kinases (ERK), JNK, and p38MAPK.
- Emphasis on practical application in mast cell research.
Main Results:
- Recent advances enable precise measurement of MAPK activity in individual mast cells.
- These methods provide insights into the dynamic regulation of mast cell signaling.
- The described techniques are robust and adaptable for various research scenarios.
Conclusions:
- Accurate assessment of MAPK activation in single mast cells is now achievable.
- These advancements are vital for dissecting complex signal transduction in mast cells.
- The reviewed methods will advance research on mast cell biology and related diseases.
Related Concept Videos
02:38Mitogens and the Cell Cycle
Mitogens and their receptors play a crucial role in controlling the progression of the cell cycle. However, the loss of mitogenic control over cell division leads to tumor formation. Therefore, mitogens and mitogen receptors play an important role in cancer research. For instance, the epidermal growth factor (EGF) - a type of mitogen and its transmembrane receptor (EGFR), decides the fate of the cell's proliferation. When EGF binds to EGFR, a member of the ErbB family of tyrosine kinase...
03:03mTOR Signaling and Cancer Progression
The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
The mTOR pathway or the...
The mTOR pathway or the...
01:19Interactions Between Signaling Pathways
Signaling cascades usually lack linearity. Multiple pathways interact and regulate one another, allowing cells to integrate and respond to diverse environmental stimuli.
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
01:07MAPK 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...
01:20The JAK-STAT Signaling Pathway
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...
01:22PI3K/mTOR/AKT Signaling Pathway
The mammalian target of rapamycin (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast, mTORC2 consists of a rapamycin-insensitive companion...