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Updated: Jun 19, 2025
Anticancer Efficacy of Photodynamic Therapy with Lung Cancer-Targeted Nanoparticles
Published on: December 1, 2016
Using Photoreactive Probes to Identify Viable Drug Targets in Non-small Cell Lung Cancer
1Department of Drug Discovery, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA. Yi.Liao@moffitt.org.
Abstract:
Recent advancements in chemoproteomics have accelerated new chemical tools for exploring protein ligandability in native biological systems. However, a large fraction of ligandable proteome in cancer cells remains poorly studied. Here, we present a practical and efficient sample processing method for liquid chromatography high-resolution-tandem mass spectrometry (HPLC-MS/MS) analysis. This method uses fully functionalized photoreactive fragment-like probes for profiling protein-ligand interactions in live cancer cells. This method adopts "on-bead" digestion in conjunction with ZipTip desalting prior sample injection to MS. By using this protocol, fragment protein interactions can be visualized using fluorescent imaging, and fragment-associated proteins can be identified via HPLC-MS/MS analysis. Approximately 16 samples would generally expect to be processed within 3 days by following this protocol.
Insights
Chemoproteomics now offers new tools to study protein interactions in cancer cells. A novel, efficient method using photoreactive probes and mass spectrometry enables rapid profiling of these interactions.
Area of Science:
- Chemical biology
- Proteomics
- Mass spectrometry
Background:
- Chemoproteomics advancements enable exploration of protein ligandability.
- The ligandable proteome in cancer cells remains largely uncharacterized.
- Novel chemical tools are needed to profile protein-ligand interactions in vivo.
Purpose of the Study:
- To develop a practical and efficient sample processing method for chemoproteomic analysis.
- To profile protein-ligand interactions in live cancer cells using photoreactive probes.
- To enable identification of fragment-associated proteins via HPLC-MS/MS.
Main Methods:
- Utilized fully functionalized photoreactive fragment-like probes.
- Employed "on-bead" digestion and ZipTip desalting for sample preparation.
- Integrated liquid chromatography-high-resolution-tandem mass spectrometry (HPLC-MS/MS) for analysis.
Main Results:
- Enabled visualization of fragment-protein interactions using fluorescent imaging.
- Facilitated identification of fragment-associated proteins.
- Demonstrated efficient processing of approximately 16 samples within 3 days.
Conclusions:
- The developed method provides a practical and efficient approach for chemoproteomic profiling.
- This protocol accelerates the study of protein-ligand interactions in cancer cells.
- The method allows for visualization and identification of protein-ligand interactions in native biological systems.
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