Enhanced Sensitivity of FRET-Based Protease Sensors by Redesign of the GFP Dimerization Interface

• Published in: Chembiochem : a European journal of chemical biology Vol. 8; no. 10; pp. 1119 - 1121
• Main Authors: Vinkenborg, Jan L., Evers, Toon H., Reulen, Sanne W. A., Meijer, E. W., Merkx, Maarten
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 09-07-2007; WILEY‐VCH Verlag
• Subjects: Bacterial Proteins - metabolism; Dimerization; Endopeptidases - chemistry; Fluorescence Resonance Energy Transfer - instrumentation; Fluorescence Resonance Energy Transfer - methods; fluorescent probes; FRET; GFP; Green Fluorescent Proteins - chemistry; Green Fluorescent Proteins - metabolism; Indicators and Reagents - pharmacology; Luminescent Proteins - metabolism; Mutation; Peptide Hydrolases - chemistry; Peptides - chemistry; proteases; protein engineering; Protein Structure, Tertiary; Sensitivity and Specificity; Time Factors
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.200700109

Close encounters. Sensor proteins based on fluorescence resonance energy transfer (FRET) often display a modest change in emission ratio upon activation. Here, we show that promoting intramolecular interactions between donor and acceptor fluorescent domains is an attractive new strategy for increasing the ratiometric change in FRET‐based protease sensors.