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A FRET-Based Biosensor for the Src N-Terminal Regulatory Element. | LitMetric

A FRET-Based Biosensor for the Src N-Terminal Regulatory Element.

Biosensors (Basel)

BioNMR Laboratory, Department of Inorganic and Organic Chemistry, Universitat de Barcelona, Baldiri Reixac, 10-12, 08028 Barcelona, Spain.

Published: February 2022

AI Article Synopsis

Article Abstract

In signaling proteins, intrinsically disordered regions often represent regulatory elements, which are sensitive to environmental effects, ligand binding, and post-translational modifications. The conformational space sampled by disordered regions can be affected by environmental stimuli and these changes trigger, vis a vis effector domain, downstream processes. The disordered nature of these regulatory elements enables signal integration and graded responses but prevents the application of classical approaches for drug screening based on the existence of a fixed three-dimensional structure. We have designed a genetically encodable biosensor for the N-terminal regulatory element of the c-Src kinase, the first discovered protooncogene and lead representative of the Src family of kinases. The biosensor is formed by two fluorescent proteins forming a FRET pair fused at the two extremes of a construct including the SH4, unique and SH3 domains of Src. An internal control is provided by an engineered proteolytic site allowing the generation of an identical mixture of the disconnected fluorophores. We show FRET variations induced by ligand binding. The biosensor has been used for a high-throughput screening of a library of 1669 compounds with seven hits confirmed by NMR.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8870054PMC
http://dx.doi.org/10.3390/bios12020096DOI Listing

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