研究目的

Investigating kinase activity in living cells remains a challenge, and usual methods are limited when one wishes to study cellular dynamic events. This review aims to address the different kinase biosensors that have been developed, how they allow specific tracking of the activity or activation of a kinase, and to give an overview of the future challenging methods to simultaneously track several biosensors in the same system.

研究成果

Genetically encoded FRET biosensors are efficient at monitoring protein activation at cellular levels when expressed in living cells. A lot of proteins are known to adopt different conformation states according to their activation, and this is why FRET or BRET biosensors are best suited for tracking their activity in space and in time in living cells. It is likely that this tool will be used intensively to study protein conformation linked to activity in the next few years.

研究不足

The biosensors rely on the endogenous kinase phosphorylating the substrate peptide, and thus, FRET variation is observed only when the kinase is particularly abundant or heavily stimulated. The sequence flanking the phosphorylation residue(s) targeted by the kinase must be known and selective for the kinase under study. These biosensors only explore the catalytic activity of the kinase toward a specific substrate at once and not the activation process of the kinase itself.