Protein and Protease Sensing by Allosteric Derepression

Abstract

Peptide motifs are crucial mediators of protein-protein interactions as well as sites of specific protease activity. The detection and characterization of these events is therefore indispensable for a detailed understanding of cellular regulation. Here, we present versatile and modular sensors that allow the user to detect protease activity and protein-peptide interactions, as well as to screen for inhibitors using chromogenic, fluorescent, or luminescent output.

Functional Display of Bioactive Peptides on the vGFP Scaffold.

“…p53 and Mdm2 have been used as a model interacting protein pair in the development of several biosensing technologies [20][21][22][23][24][25] . We have previously described a biosensor comprising b-lactamase coupled to an inhibitory protein (BLIP) in cis via a linker containing the M2 peptide sequence 26,27 . Upon binding to Mdm2, the normally disordered M2 peptide adopted an a-helical conformation, effectively shortening linker length and allosterically displacing BLIP and restoring b-lactamase activity.…”