Premium
Cover Picture: Enhanced Sensitivity of FRET‐Based Protease Sensors by Redesign of the GFP Dimerization Interface (ChemBioChem 10/2007)
Author(s) -
Vinkenborg Jan L.,
Evers Toon H.,
Reulen Sanne W. A.,
Meijer E. W.,
Merkx Maarten
Publication year - 2007
Publication title -
chembiochem
Language(s) - English
Resource type - Reports
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200790029
Subject(s) - förster resonance energy transfer , intramolecular force , linker , green fluorescent protein , fluorescence , biophysics , protease , chemistry , cyan , protein engineering , cleavage (geology) , yellow fluorescent protein , peptide , conformational change , photochemistry , materials science , stereochemistry , biochemistry , biology , physics , computer science , quantum mechanics , fracture (geology) , optics , composite material , gene , enzyme , operating system
The cover picture shows a new strategy to increase the sensitivity of protease sensors based on fluorescence resonance energy transfer between enhanced cyan and yellow fluorescent protein domains (ECFP and EYFP). Introduction of two mutations (S208F and V224L) at the dimerization interface of the two fluorescent domains induces intramolecular complex formation and results in a strong increase in energy transfer efficiency. Proteolytic cleavage of the peptide linker disrupts the intramolecular interaction yielding an impressive 16‐fold change in emission ratio. These results demonstrate that promoting intramolecular domain interactions can be an attractive strategy to improve the modest emission ratio changes often observed for FRET‐based sensor proteins. For further information see the Communication by M. Merkx, et al. on p. 1119 ff.