Characterization of Split Fluorescent Protein Variants and Quantitative Analyses of Their Self-Assembly Process.

ABSTRACT: Many biotechniques use complementary split-fluorescent protein (sFPs) fragments to visualize protein-protein interactions, image cells by ensemble or single molecule fluorescence microscopy, or assemble nanomaterials and protein superstructures. Yet, the reassembly mechanisms of sFPs, including fragment binding rates, folding, chromophore maturation and overall photophysics remain poorly characterized. Here, we evolved asymmetric and self-complementing green, yellow and cyan sFPs together with their full-length equivalents (flFPs) and described their biochemical and photophysical properties in vitro and in cells. While re-assembled sFPs have spectral properties similar to flFPs, they display slightly reduced quantum yields and fluorescence lifetimes due to a less sturdy β-barrel structure.

SUBMITTER: Koker T 

PROVIDER: S-EPMC5871787 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature