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A cell permeable NPE caged ADP-ribose for studying TRPM2
Peilin Yu1, Qian Wang, Li-He Zhang
1Department of Physiology, University of Hong Kong, Hong Kong, China.
Plos One
|December 14, 2012
Summary
Researchers developed a new caged adenosine diphosphate ribose (ADPR) called NPE-ADPR to study the TRPM2 channel. This tool effectively triggers cation influx, aiding in the investigation of TRPM2-mediated cellular processes.
Area of Science:
- Ion channel research
- Cellular signaling
- Biochemistry
Background:
- Transient potential receptor melastatin-2 (TRPM2) is a calcium-permeable cation channel.
- TRPM2 is primarily activated by intracellular adenosine diphosphate ribose (ADPR).
- Studying TRPM2 function in intact cells requires specific tools to control ADPR levels.
Purpose of the Study:
- To synthesize and validate a novel caged ADPR, NPE-ADPR.
- To investigate the utility of NPE-ADPR in studying TRPM2-mediated cation influx in intact cells.
- To characterize the conditions affecting NPE-ADPR-induced cation entry.
Main Methods:
- Synthesis of 1-(2-nitrophenyl)ethyl caged ADPR (NPE-ADPR).
- Application of NPE-ADPR to intact human Jurkat T-lymphocytes and HEK 293 cells overexpressing TRPM2.
- Measurement of cation influx (Ca2+, Mg2+, Zn2+) using various techniques.
- Utilized TRPM2 inhibitors and knockdown, temperature, pH, and extracellular ion concentration manipulations.
Main Results:
- Photolysis of NPE-ADPR efficiently stimulated concentration-dependent Ca2+, Mg2+, and Zn2+ influx in Jurkat T-cells and HEK 293 cells overexpressing TRPM2.
- TRPM2 inhibition or knockdown significantly reduced NPE-ADPR-induced cation influx.
- NPE-ADPR-induced cation entry was modulated by temperature and pH, and dependent on extracellular Ca2+ for Mg2+ and Zn2+ influx.
Conclusions:
- NPE-ADPR is a cell-permeable ADPR analog.
- NPE-ADPR is a valuable tool for studying TRPM2-mediated cation entry in intact cellular systems.
- This method provides a controlled way to activate TRPM2 channels for research purposes.
