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Protein Dynamics in Living Cells

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.
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Updated: Feb 19, 2026

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

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Published on: January 16, 2019

Development of a Protease Biosensor Based on a Dimerization-Dependent Red Fluorescent Protein.

Aaron C Mitchell¹, Spencer C Alford¹, Sean A Hunter²

¹Department of Bioengineering, Stanford University , Stanford, California 94305, United States.

ACS Chemical Biology

|November 11, 2017

Summary

Researchers developed a novel matriptase biosensor using a red fluorescent protein system. This tool accurately detects matriptase activity and inhibition, aiding in cancer and osteoarthritis research.

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Published on: January 16, 2019

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Determining Membrane Protein Topology Using Fluorescence Protease Protection FPP

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Published on: April 20, 2015

Area of Science:

Biochemistry
Molecular Biology
Biotechnology

Background:

Matriptase protease activity is linked to aggressive tumor growth, metastasis, and osteoarthritis.
Accurate detection and quantification methods for matriptase activity and inhibition are needed.

Purpose of the Study:

To develop a novel matriptase-sensitive protein biosensor for detecting and quantifying matriptase activity and inhibition.
To create a versatile platform for studying other proteases.

Main Methods:

Utilized a dimerization-dependent red fluorescent protein (ddRFP) reporter system.
Engineered biosensors with protease-labile linkers between protein domains.
Characterized linker cleavage, matriptase activity, and specificity of various ddRFP constructs.

Main Results:

Developed a highly efficient and specific matriptase biosensor (B4) with an RSKLRVGGH linker.
Demonstrated detection of matriptase inhibition with a 5-fold dynamic range.
Successfully detected matriptase activity in human cancer cell lines.

Conclusions:

The developed ddRFP-based biosensor is effective for detecting matriptase activity and inhibition.
This platform offers a valuable tool for cancer and osteoarthritis research.
The strategy can be adapted for biosensors targeting other proteases.