Reduced Fluorescent Protein Switching Fatigue by Binding-Induced Emissive State Stabilization.

ABSTRACT: Reversibly switchable fluorescent proteins (RSFPs) enable advanced fluorescence imaging, though the performance of this imaging crucially depends on the properties of the labels. We report on the use of an existing small binding peptide, named Enhancer, to modulate the spectroscopic properties of the recently developed rsGreen series of RSFPs. Fusion constructs of Enhancer with rsGreen1 and rsGreenF revealed an increased molecular brightness and pH stability, although expression in living E. coli or HeLa cells resulted in a decrease of the overall emission. Surprisingly, Enhancer binding also increased off-switching speed and resistance to switching fatigue. Further investigation suggested that the RSFPs can interconvert between fast- and slow-switching emissive states, with the ove

SUBMITTER: Roebroek T 

PROVIDER: S-EPMC5618663 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature