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A rapid and sensitive assay for histone acetyl-transferase activity
S Ait-Si-Ali1, S Ramirez, P Robin
1Laboratoire Oncogénèse, Différenciation et Transduction du Signal, CNRS UPR 9079, IFC-01, 7 rue Guy Moquet, 94801 Villejuif, France.
Nucleic Acids Research
|August 1, 1998
Summary
We developed a sensitive and rapid enzymatic assay to measure histone acetyl-transferase (HAT) activity. This method quantifies HATs by measuring radioactivity after acetylating a histone H4 peptide substrate.
Area of Science:
- Biochemistry
- Molecular Biology
- Enzymology
Background:
- Histone acetyl-transferases (HATs) play a crucial role in gene transcription regulation.
- Accurate quantification of HAT enzymatic activity is essential for understanding epigenetic mechanisms.
Purpose of the Study:
- To design and validate a novel enzymatic assay for quantifying HAT activity.
- To provide a sensitive, rapid, and convenient method for HAT activity measurement.
Main Methods:
- A biotinylated peptide substrate representing the first 24 amino acids of histone H4 was synthesized.
- Substrate acetylation was performed using [14C]acetyl-CoA.
- Purification of acetylated peptide was achieved using streptavidin beads.
- Associated radioactivity was measured to quantify HAT activity.
Main Results:
- The developed assay effectively quantifies HAT enzymatic activity.
- The assay demonstrates high sensitivity, speed, and convenience.
Conclusions:
- This novel enzymatic assay is a valuable tool for studying HAT function.
- The assay facilitates research into transcription regulation and epigenetic modifications.