Related Experiment Video
Updated: Aug 6, 2026
07:59
Temporal Quantification of MAPK Induced Expression in Single Yeast Cells
Published on: October 4, 2013
Expression and characterization of MAP kinases in bacteria
Charles J Heise1, Melanie H Cobb
1Department of Pharmacology, The University of Texas Southwestern Medical Center at Dallas, 6001 Forest Park Road, Dallas, TX 75390-9041, USA.
Methods (San Diego, Calif.)
|August 16, 2006
Summary
Mitogen-activated protein kinases (MAPKs) are crucial in eukaryotic cell signaling. This study details methods to phosphorylate or thiophosphorylate ERK2 in vitro, aiding MAPK network research.
Area of Science:
- Cellular Biology
- Molecular Biology
- Biochemistry
Background:
- Mitogen-activated protein kinases (MAPKs) are essential signal transducers in eukaryotes.
- MAPKs, including Extracellular-signal regulated kinases 1 and 2 (ERK1/2), are activated via kinase cascades (MAP3Ks, MAP2Ks).
- MAPK activity is tightly regulated by phosphorylation and dephosphorylation.
Purpose of the Study:
- To describe a method for in vitro phosphorylation and thiophosphorylation of ERK2.
- To provide a tool for investigating mitogen-activated protein kinase signaling pathways.
Main Methods:
- In vitro phosphorylation of ERK2.
- In vitro thiophosphorylation of ERK2.
Main Results:
- Successful phosphorylation of ERK2 in vitro.
- Successful thiophosphorylation of ERK2 in vitro.
Conclusions:
- The described in vitro phosphorylation and thiophosphorylation methods are valuable for studying MAPK signaling.
- These techniques enable further investigation into the complex MAPK signaling networks.
Related Concept Videos
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...
01:42Microtubule Associated Proteins (MAPs)
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...
01:29Cytoskeletal Proteins in Bacteria
Bacterial cells were initially considered simple, randomly organized structures lacking a cytoskeleton. However, the discovery of cytoskeleton homologs in bacteria led to the change of this opinion. Bacterial cytoskeletal filaments regulate the cell shape, cell polarity, cell division, and partitioning of plasmids during cell division. It was later discovered that bacterial cytoskeletal proteins, mainly actin and tubulin homologs, are diverse compared to their eukaryotic counterparts. On the...
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...
01:16Calmodulin-dependent Signaling
Calmodulin (CaM) is a calcium-binding protein in eukaryotes that controls various calcium-regulated cellular processes. It has four calcium-binding sites that bind calcium to form the calcium-calmodulin ( Ca2+-CaM) complex. GPCR stimulation increases the calcium levels in the cells that bind to CaM and induces a conformational change.
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...
01:28Regulation of Bacterial Virulence
Pathogenic bacteria employ a range of regulatory mechanisms to modulate the expression of virulence genes in response to environmental and host-derived signals. These mechanisms ensure that virulence factors are expressed only under favorable conditions, thereby optimizing infection and survival strategies.Mechanisms of Virulence RegulationKey regulatory strategies include:Two-Component Systems: These consist of a membrane-bound sensor kinase and a cytoplasmic response regulator. Environmental...