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Measurement of Force-Sensitive Protein Dynamics in Living Cells Using a Combination of Fluorescent Techniques
Published on: November 2, 2018
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Quantifying cadherin mechanotransduction machinery assembly/disassembly dynamics using fluorescence covariance
Pavan Vedula1, Lissette A Cruz1, Natasha Gutierrez1
1Department of Biological Sciences, Rutgers, The State University of New Jersey, Newark, New Jersey, USA.
Scientific Reports
|July 1, 2016
Summary
This study quantifies multi-protein adherens junction complex assembly in living cells using Fluorescence Covariance Analysis. The findings reveal key stages of complex formation and maturation during epithelial tissue development.
Area of Science:
- Cell Biology
- Biophysics
- Biochemistry
Background:
- Understanding multi-molecular complex assembly in cellular compartments is vital for cell function.
- Current methods for studying complex dynamics have limitations in quantifying in situ assembly.
Purpose of the Study:
- To validate and quantify multi-protein adherens junction complex assembly in situ.
- To analyze the dynamics of adherens junction complex assembly during epithelial monolayer formation.
Main Methods:
- Utilized light microscopy and Fluorescence Covariance Analysis (FCA).
- Employed fluorescently-labeled protein pairs to track complex assembly.
- Applied FCA to live cells expressing tagged adherens junction proteins.
Main Results:
- Quantified stages of adherens junction complex assembly, from perinuclear localization to cell-cell contact zones.
- Demonstrated acto-myosin tension-mediated anchoring and maturation of the complex.
- Measured adherens junction complex assembly dynamics during epithelial monolayer formation.
Conclusions:
- Fluorescence Covariance Analysis is a validated method for quantifying in situ multi-protein complex assembly.
- Adherens junction complex assembly is a dynamic process involving sequential steps and cytoskeletal interactions.
- This approach provides quantitative insights into epithelial tissue development and regulation.
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