Highly photostable fluorescent labeling of proteins in live cells using exchangeable coiled coils heterodimerization.

ABSTRACT: Fluorescent proteins are commonly used to label target proteins in live cells. However, the conventional approach based on covalent fusion of targeted proteins with fluorescent protein probes is limited by the slow rate of fluorophore maturation and irretrievable loss of fluorescence due to photobleaching. Here, we report a genetically encoded protein labeling system utilizing transient interactions of small, 21-28 residues-long helical protein tags (K/E coils, KEC). In this system, a protein of interest, covalently tagged with a single coil, is visualized through binding to a cytoplasmic fluorescent protein carrying a complementary coil. The reversible heterodimerization of KECs, whose affinity can be tuned in a broad concentration range from nanomolar to micromolar, allows continuous exc

SUBMITTER: Perfilov MM 

PROVIDER: S-EPMC7329588 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature