Related Experiment Video

Updated: Mar 18, 2026

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy
10:41

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Published on: June 7, 2019

9.1K

In-situ coupling between kinase activities and protein dynamics within single focal adhesions

Yiqian Wu1, Kaiwen Zhang1, Jihye Seong2

  • 1Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093, USA.

Scientific Reports
|July 8, 2016
PubMed

Insights

This study reveals how kinase activation and focal adhesion dynamics are coordinated at single sites in cancer cells. Tightly coupled events promote focal adhesion growth, while less coupled events lead to disassembly, highlighting Src and FAK interplay.

Area of Science:

  • Cell Biology
  • Cancer Research
  • Biophysics

Background:

  • Cell adhesion and focal adhesion (FA) dynamics are critical in cancer metastasis.
  • The temporal coordination of oncogenic kinase activation and FA regulation at single sites remains poorly understood.

Purpose of the Study:

  • To investigate the temporal interplay between kinase activity and FA dynamics at the subcellular level.
  • To elucidate the roles of Src and focal adhesion kinase (FAK) in regulating the life cycle of single FAs.

Main Methods:

  • Utilized fluorescence resonance energy transfer (FRET)-based biosensors targeted to subcellular FAs.
  • Employed single FA tracking and cross-correlation analysis to quantify dynamic coupling.
  • Integrated computational analysis to interpret molecular events.

Main Results:

  • Demonstrated a sequential correlation between kinase activation and FA assembly, with FA assembly and Src activation preceding FAK activation.
  • Showed that the temporal coupling between kinase activation and FA assembly predicts FA fate (maturation or disassembly).
  • Observed that during FA disassembly, kinase activation precedes disassembly, with FAK activation occurring before Src activation.

Conclusions:

  • Revealed intricate interplays between Src and FAK in regulating the dynamic life of single FAs.
  • Established a link between the temporal coupling of molecular events and the functional outcome of FAs.
  • Provided insights into the spatiotemporal regulation of cell adhesion in cancer metastasis.

More Related Videos

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET13:38

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET

Published on: January 27, 2012

14.7K
Measurement of Force-Sensitive Protein Dynamics in Living Cells Using a Combination of Fluorescent Techniques08:28

Measurement of Force-Sensitive Protein Dynamics in Living Cells Using a Combination of Fluorescent Techniques

Published on: November 2, 2018

8.9K

Related Concept Videos

Intracellular Signaling Affects Focal Adhesions01:17

Intracellular Signaling Affects Focal Adhesions

Integrins act both as extracellular input receivers and as intracellular processing activators. As their name suggests, integrins are entirely integrated into the membrane structure. Their hydrophobic membrane-spanning regions interact with the phospholipid bilayer's hydrophobic region. These membrane receptors provide extracellular attachment sites for effectors like hormones and growth factors. They activate intracellular response cascades when their effectors are bound and active.
Some...
3.8K
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.8K
Cytoskeletal Coordination in Cell Migration01:32

Cytoskeletal Coordination in Cell Migration

A migrating cell changes its shape during the cyclic events of attachment and detachment from the substratum and repositions the cell organelles correspondingly. These complex events are orchestrated by the dynamic cytoskeletal network comprising actin filaments, intermediate filaments, and microtubules. Cytoskeletal crosstalk — the direct and indirect communication between the different components — is crucial for this coordination. Direct communication involves various linker...
5.7K
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

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...
19.0K
Tension Response at Adherens Junctions01:26

Tension Response at Adherens Junctions

The adherens junctions that anchor cells together are multi-protein complexes that dynamically adapt to mechanical stimuli such as tensile forces and shear stress. Mechanosensory proteins in these junctions can sense such mechanical stimuli and undergo a shift in their conformation, resulting in an altered function — a process called mechanotransduction.
α-Catenin as a Mechanosensory Protein
The α-catenin of adherens junctions is an allosteric protein with three VH (vinculin...
4.1K
Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
7.0K