protein usually needs to "work" through changes in the conformation and . However, the existing tools for tracking protein conformational changes in living cells are still lacking, and in many cases they cause great interference to the system or are not scalable (1, 2).
Researchers such as Klaus Hahn and Timothy Elston at the University of North Carolina at Chapel Hill have developed a new system with strong scalability and low system interference to study protein conformational changes in living cells (2).
The core idea of this system is to add a small tag (only 7 amino acids are used in the article) at the position where the target protein is exposed and hidden according to the conformation. This small tag can specifically bind to the fluorescently labeled reporter protein. , thereby converting protein conformational changes into fluorescent signals. The researchers further combined FRET to improve the specificity of the system and studied the dynamic changes in the spatiotemporal activity of , a kinase (Src) (2).
Live cells track protein activity changes (2)
Combined with FRET to further improve system specificity (2)
Researchers believe that this system can be flexibly adjusted to study previously inoperable protein conformational changes in living cells (2).
This work was published in Cell(2) on October 11, 2021.
Comments:
tags are relatively small and have strong scalability, which are important advantages; however, this system has very high requirements for the expression of the reporter protein, and is very particular about the insertion position of the "tag", which requires very detailed optimization to achieve the ideal effect.
https://www.cell.com/cell/fulltext/S0092-8674(21)01109-0
Corresponding author introduction:
https://www.med.unc.edu/pharm/directory/klaus-m-hahn-phd /
https://www.med.unc.edu/pharm/directory/timothy-elston/
Reference:
1. J.-E. EA, J. TM, FRET imaging. Nat.Biotechnol. 21, 1387–1395 (2003).
2. B. Liu et al., Biosensors based on peptide exposure show single molecule conformations in live cells. Cell.0 (2021), doi:10.1016/J.CELL.2021.09.026.
Original link:
https ://www.cell.com/cell/fulltext/S0092-8674(21)01109-0
