Related Experiment Video
Updated: May 11, 2026
08:37
Fluorescence-based Monitoring of PAD4 Activity via a Pro-fluorescence Substrate Analog
Published on: November 5, 2014
Facile fluorescence-based detection of PAD4-mediated citrullination
Erin Wildeman1, Marcos M Pires
1Department of Chemistry, Lehigh University, 6 E. Packer Ave., Bethlehem, PA 18015, USA.
Summary
We developed a simple, fluorescence-based assay to detect Protein Arginine Deiminase 4 (PAD4) activity. This assay can identify PAD4
Area of Science:
- Biochemistry
- Molecular Biology
- Enzymology
Background:
- Post-translational modifications of histone proteins are crucial for cellular homeostasis and disease.
- Protein arginine deiminase 4 (PAD4) enzyme activity is linked to cancer and autoimmune diseases.
- Accurate detection of PAD4 activity is essential for understanding its role in disease.
Purpose of the Study:
- To develop a facile, fluorescence-based assay for detecting Protein Arginine Deiminase 4 (PAD4) activity.
- To provide a rapid and reliable method for monitoring PAD4-mediated citrullination.
- To enable the study of PAD4 in cellular homeostasis and disease states.
Main Methods:
- Developed a fluorescence-based assay utilizing trypsin's substrate specificity.
- Monitored the citrullination reaction by detecting the conversion of arginine to citrulline.
- Assessed assay performance with a known PAD4 inhibitor.
Main Results:
- The assay successfully detects PAD4 activity through citrullination.
- The assay is rapid, uses readily available reagents, and is sensitive to PAD4 inhibition.
- Demonstrated the conversion of positively charged arginine to neutral citrulline.
Conclusions:
- A novel, facile, fluorescence-based assay for PAD4 activity has been established.
- This assay offers a sensitive and efficient method for studying PAD4 function.
- The assay has potential applications in disease research and drug discovery.
