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A Proteoliposome-Based Efflux Assay to Determine Single-molecule Properties of Cl- Channels and Transporters
Published on: April 20, 2015
10.3K
Synthetic probe development for measuring single or few-cell activity and efflux
Alison Lui1, Jeffrey Wang1, Linda Chio1
1Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA, United States.
Methods in Enzymology
|November 1, 2019
Summary
This study introduces polymer-functionalized carbon nanotube sensor arrays for imaging single-cell protein secretion. This technology reveals spatiotemporal heterogeneity in protein release, crucial for understanding cellular communication and function.
Area of Science:
- Biophysics
- Cell Biology
- Nanotechnology
Background:
- Understanding cellular communication requires analyzing protein secretion from individual cells.
- Heterogeneous protein secretion impacts overall physiological and signaling processes.
- Spatiotemporal resolution is key to linking cellular structure and function in secretion.
Purpose of the Study:
- To present protocols for creating and utilizing polymer-functionalized single-walled carbon nanotube (SWCNT) sensor arrays.
- To demonstrate the capability of SWCNT sensor arrays in imaging single-cell protein secretion.
- To explore the potential of this platform for studying diverse analyte effluxes.
Main Methods:
- Development of polymer-functionalized SWCNT sensor arrays.
- Protocols for sensor creation, array preparation, and imaging secreted proteins.
- Application in both prokaryotic and mammalian cell systems.
Main Results:
- Successful microscopic imaging of secreted protein footprints at the single-cell level.
- Demonstration of spatiotemporal heterogeneity in protein secretion.
- Proof-of-concept imaging of secreted RAP1 and HIV-1 integrase proteins.
Conclusions:
- SWCNT sensor arrays provide a powerful tool for studying single-cell secretomes.
- The platform enables detailed analysis of spatiotemporal protein secretion dynamics.
- Future perspectives include broader sensor applications and development for various analytes.
