Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

5.8K
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...
5.8K
MAPK Signaling Cascades01:07

MAPK Signaling Cascades

8.8K
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...
8.8K
Receptor Tyrosine Kinases01:26

Receptor Tyrosine Kinases

19.8K
Receptor tyrosine kinases or RTKs are membrane-bound receptors that phosphorylate specific tyrosine on protein substrates. RTKs regulate cellular growth, differentiation, survival, and migration. They contain an extracellular ligand binding domain, a transmembrane domain, and a cytosolic tail with intrinsic kinase activity. Several extracellular signaling molecules activate RTKs in one or more ways and relay the signal downstream. Ligands such as platelet-derived growth factor (PDGF) or...
19.8K
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

18.7K
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...
18.7K
The JAK-STAT Signaling Pathway01:20

The JAK-STAT Signaling Pathway

13.3K
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...
13.3K
Interactions Between Signaling Pathways01:19

Interactions Between Signaling Pathways

7.4K
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...
7.4K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Targeting p130Cas- and microtubule-dependent MYC regulation sensitizes pancreatic cancer to ERK MAPK inhibition.

Cell reports·2026
Same author

Defining the MYC-regulated transcriptome and kinome that support KRAS- and ERK-dependent growth of pancreatic cancer.

Science signaling·2025
Same author

Local optogenetic control of genome editing and tumorigenesis <i>in vivo</i> using wireless implantable optoelectronics.

bioRxiv : the preprint server for biology·2025
Same author

Spatiotemporal Control of Protein Activity through Optogenetic Allosteric Regulation.

Journal of visualized experiments : JoVE·2024
Same author

Development and Application of Rapamycin-regulated Tyrosine Phosphatases.

Journal of visualized experiments : JoVE·2024
Same author

Determining the ERK-regulated phosphoproteome driving KRAS-mutant cancer.

Science (New York, N.Y.)·2024

Related Experiment Video

Updated: Feb 26, 2026

Author Spotlight: Developing Tools to Tune the Activity of Tyrosine Phosphatases
06:56

Author Spotlight: Developing Tools to Tune the Activity of Tyrosine Phosphatases

Published on: September 6, 2024

843

Dissecting Kinase Effector Signaling Using the RapRTAP Methodology.

Anne-Marie Ray1, Jennifer E Klomp1, Kerrie B Collins1

  • 1Department of Pharmacology, University of Illinois at Chicago, Chicago, IL, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 22, 2017
PubMed
Summary

Researchers developed a novel tool to selectively activate kinases within specific protein complexes in living cells. This method allows precise control over kinase signaling, enabling detailed study of cellular processes and disease mechanisms.

Keywords:
FKBP12KinasePhosphorylationRapamycinTargeted signaling pathway

More Related Videos

A Semi-Quantitative Drug Affinity Responsive Target Stability DARTS assay for studying Rapamycin/mTOR interaction
05:28

A Semi-Quantitative Drug Affinity Responsive Target Stability DARTS assay for studying Rapamycin/mTOR interaction

Published on: August 27, 2019

17.8K
Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

8.6K

Related Experiment Videos

Last Updated: Feb 26, 2026

Author Spotlight: Developing Tools to Tune the Activity of Tyrosine Phosphatases
06:56

Author Spotlight: Developing Tools to Tune the Activity of Tyrosine Phosphatases

Published on: September 6, 2024

843
A Semi-Quantitative Drug Affinity Responsive Target Stability DARTS assay for studying Rapamycin/mTOR interaction
05:28

A Semi-Quantitative Drug Affinity Responsive Target Stability DARTS assay for studying Rapamycin/mTOR interaction

Published on: August 27, 2019

17.8K
Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

8.6K

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Kinases regulate diverse cellular functions, and understanding their signaling pathways is crucial for cell biology.
  • Targeting specific kinase substrates remains a significant challenge in cell signaling research.

Purpose of the Study:

  • To present a novel tool for the selective activation of kinases within specific protein complexes in living cells.
  • To demonstrate the application of this technology in dissecting kinase-mediated signaling pathways.

Main Methods:

  • Utilized a rapamycin-inducible system for kinase activation.
  • Coupled kinase activation to FKBP12-binding domain (FRB) tagged proteins for targeted localization.
  • Applied the method to target Src kinase to p130Cas and FAK proteins.

Main Results:

  • Successfully demonstrated selective activation of Src kinase in specific protein complexes.
  • Discriminated distinct cell morphodynamic changes downstream of Src targeting to p130Cas versus FAK.
  • Provided a method to dissect kinase-specific signaling outputs.

Conclusions:

  • The developed tool enables precise control over kinase activity in targeted cellular contexts.
  • This technology facilitates the elucidation of kinase-substrate interactions and downstream signaling.
  • Offers a valuable approach for studying physiological and pathological cellular processes involving kinases.