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Updated: Mar 29, 2026
Author Spotlight: Enhanced Histone PTM Isomer Identification Through LC-TIMS-ToF MS/MS and PASEF
Published on: January 12, 2024
Multiplex detection of histone-modifying enzymes by total internal reflection fluorescence-based single-molecule
Fei Ma1, Meng Liu2, Zi-yue Wang2
1College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Shandong Provincial Key Laboratory of Clean Production of Fine Chemicals, Shandong Normal University, Jinan 250014, China. cyzhang@sdnu.edu.cn and Single-Molecule Detection and Imaging Laboratory, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China.
We developed a sensitive method for detecting histone-modifying enzymes (HMEs) using fluorescence and single-molecule detection. This technique enables efficient screening of HME inhibitors.
Area of Science:
- Biochemistry
- Molecular Biology
- Enzymology
Background:
- Histone-modifying enzymes (HMEs) play crucial roles in epigenetic regulation.
- Accurate detection of HMEs is essential for understanding gene expression and disease.
- Existing methods for HME detection may lack sensitivity or multiplexing capabilities.
Purpose of the Study:
- To develop a novel, highly sensitive, and selective method for the multiplex detection of histone-modifying enzymes (HMEs).
- To enable the screening of potential inhibitors for HMEs.
Main Methods:
- Integration of antibody-based fluorescence labeling with total internal reflection fluorescence (TIRF)-based single-molecule detection.
- Utilized specific antibodies for labeling target HMEs.
- Employed TIRF microscopy for high-sensitivity, single-molecule level detection.
Main Results:
- Achieved excellent specificity and high sensitivity in HME detection.
- Demonstrated a detection limit of 21 pM for histone acetyltransferase GcN5.
- Achieved a detection limit of 12 pM for histone methyltransferase G9a.
- Successfully applied the method for screening HME inhibitors.
Conclusions:
- The developed method offers a sensitive and selective approach for multiplex HME detection.
- This technique is valuable for biochemical assays and drug discovery, particularly for HME inhibitors.
- The single-molecule detection capability provides a powerful tool for studying enzyme kinetics and interactions.
