Profiling Ligand-Induced Changes in Nuclear Localization Using Proximity Labeling-Coupled Chemoproteomics.

Area of Science:

• Cell Biology
• Proteomics
• Drug Discovery

Background:

• Nuclear proteins regulate essential cellular processes like transcription and chromatin structure.
• Dysregulation of nuclear proteins is linked to diseases, including cancer.
• Targeting nuclear proteins with small molecules is crucial for therapeutic development.

Purpose of the Study:

• To develop a method for specifically analyzing the nuclear proteome.
• To identify ligandable sites within nuclear proteins for covalent targeting.
• To monitor changes in nuclear protein localization and chromatin association.

Main Methods:

• Coupling proximity labeling with histone-TurboID and chemoproteomics.
• Generating cell lines expressing histone-TurboID.
• Utilizing tandem mass tag (TMT)-based quantitative proteomics.

Main Results:

• A platform to specifically investigate the nuclear proteome was established.
• The method enables identification of ligandable sites in nuclear proteins.
• Changes in protein localization and chromatin association upon covalent ligand exposure can be monitored.

Conclusions:

• This methodology offers a streamlined approach for discovering covalent ligands targeting nuclear proteins.
• The technique facilitates the identification of novel therapeutic strategies for nuclear protein-related diseases.
• It provides insights into nuclear protein dynamics and drug interactions.