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Genetically encoded pH sensor for tracking surface proteins through endocytosis.

Anmol Grover¹, Brigitte F Schmidt, Russell D Salter

¹Department of Biological Sciences and Molecular Biosensor and Imaging Center, Carnegie Mellon University, Pittsburgh, PA 15213, USA.

Angewandte Chemie (International Ed. in English)

|March 31, 2012

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A novel tandem dye, Traffic Cam, acts as a ratiometric pH indicator for cell surfaces. This tool tracks protein trafficking during endocytosis and receptor internalization, aiding in cellular process analysis.

Area of Science:

Biochemistry
Cell Biology
Molecular Imaging

Background:

Understanding protein trafficking and receptor internalization is crucial for cellular signaling.
Existing methods for tracking these processes can be limited in scope or resolution.

Purpose of the Study:

To develop and validate a novel ratiometric pH indicator for monitoring cell surface events and endocytosis.
To utilize this indicator to study the internalization dynamics of specific receptors.

Main Methods:

Preparation of a tandem dye (Traffic Cam) comprising a FRET acceptor and a fluorogenic donor.
Application of the dye as a cell surface ratiometric pH indicator.
Monitoring protein trafficking during endocytosis and agonist-dependent receptor internalization.

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Main Results:

Traffic Cam successfully functions as a ratiometric pH indicator on the cell surface.
The sensor effectively tracks protein trafficking following internalization.
Agonist-dependent internalization of β(2)-adrenergic receptors was analyzed using this system.

Conclusions:

The Traffic Cam sensor provides a versatile tool for studying cell surface dynamics and endocytosis.
This indicator facilitates the analysis of receptor internalization pathways.
The technology shows potential for investigating antigen transfer between cells.