Genetically Encoded FRET Biosensor for Visualizing EphA4 Activity in Different Compartments of the Plasma Membrane.

Abstract

The EphA4 receptor tyrosine kinase is well-known for its pivotal role in development, cancer progression, and neurological disorders. However, how EphA4 kinase activity is regulated in time and space still remains unclear. To visualize EphA4 activity in different membrane microdomains, we developed a sensitive EphA4 biosensor based on Förster resonance energy transfer (FRET), and targeted it in or outside raft-like microdomains in the plasma membrane. We showed that our biosensor can produce a robust and specific FRET response upon EphA4 activation, both in vitro and in live cells. Interestingly, we observed stronger FRET responses for the non-raft targeting biosensor than for the raft targeting biosensor, suggesting that stronger EphA4 activation may occur in non-raft regions. Further investigations revealed the importance of the actin cytoskeleton in suppressing EphA4 activity in raft-like microdomains. Therefore, our FRET-based EphA4 biosensor could serve as a powerful tool to visualize and investigate EphA4 activation and signaling in specific subcellular compartments of single live cells.

• Authors: Yijia Pan, Shaoying Lu, Lei Lei, Ilaria Lamberto, Yi Wang, Elena B Pasquale, Yingxiao Wang
• Journal: ACS sensors (ACS Sens) Vol. 4 Issue 2 Pg. 294-300 (02 22 2019) ISSN: 2379-3694 [Electronic] United States
• PMID: 30608127 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Receptor, EphA4

Topics

• Actin Cytoskeleton (metabolism)
• Biosensing Techniques (methods)
• Cell Membrane (metabolism)
• Fluorescence Resonance Energy Transfer
• HEK293 Cells
• HeLa Cells
• Humans
• Receptor, EphA4 (metabolism)