University of Pittsburgh

Superecliptic pHluorin (SEP)-PTH Receptor (SEP-PTHR) Stable Cells

The technology employs stable cell lines engineered to express a parathyroid hormone receptor (PTHR) with an integrated Superecliptic pHluorin (SEP) tag positioned in the extracellular domain. This design leverages the pH-sensitive properties of the SEP tag, which emits a bright fluorescent signal at neutral pH when the receptor is on the cell surface and is quenched in acidic compartments during receptor internalization. Consequently, researchers can monitor receptor localization, trafficking, and recycling in real time, enabling a dynamic study of PTHR regulation and its cellular pathways.

Description

This approach is differentiated by its unique ability to visualize receptor dynamics with unmatched clarity. Unlike previous methods, the SEP-tagged PTHR provides a direct readout of receptor distribution, marking a significant advancement in live-cell imaging technologies. Its capability to distinguish between plasma membrane-bound and internalized receptors allows for advanced investigation of signaling mechanisms. Additionally, as the first application of SEP-tagging to PTHR, this technology fills a critical gap in research tools for studying bone metabolism and calcium homeostasis, offering new avenues for drug discovery and the development of therapeutic strategies.

Applications

- Drug discovery platform
- Realtime trafficking assay
- Bone metabolism research
- GPCR internalization analysis

Advantages

- Real-time visualization of PTH receptor localization using a pH-sensitive fluorescent tag.
- Clear distinction between membrane-bound and internalized receptors due to pH-dependent fluorescence.
- Dynamic monitoring of receptor trafficking, internalization, and recycling in live cells.
- Enhanced insights into PTHR regulation, aiding basic research and drug discovery.
- Novel tool that fills a critical gap in studying bone metabolism and related disorders.

IP Status

Research Tool