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Structure-guided design of a reversible fluorogenic reporter of protein-protein interactions.

ABSTRACT: A reversible green fluorogenic protein-fragment complementation assay was developed based on the crystal structure of UnaG, a recently discovered fluorescent protein. In living mammalian cells, the nonfluorescent fragments complemented and rapidly became fluorescent upon rapamycin-induced FKBP and Frb protein interaction, and lost fluorescence when the protein interaction was inhibited. This reversible fluorogenic reporter, named uPPI [UnaG-based protein-protein interaction (PPI) reporter], uses bilirubin (BR) as the chromophore and requires no exogenous cofactor. BR is an endogenous molecule in mammalian cells and is not fluorescent by itself. uPPI may have many potential applications in visualizing spatiotemporal dynamics of PPIs.

SUBMITTER: To TL

PROVIDER: S-EPMC4815423 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature

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Structure-guided design of a reversible fluorogenic reporter of protein-protein interactions.

To Tsz-Leung TL Zhang Qiang Q Shu Xiaokun X

Protein science : a publication of the Protein Society 20160109 3

A reversible green fluorogenic protein-fragment complementation assay was developed based on the crystal structure of UnaG, a recently discovered fluorescent protein. In living mammalian cells, the nonfluorescent fragments complemented and rapidly became fluorescent upon rapamycin-induced FKBP and Frb protein interaction, and lost fluorescence when the protein interaction was inhibited. This reversible fluorogenic reporter, named uPPI [UnaG-based protein-protein interaction (PPI) reporter], uses ...[more]

PMID: 26690964

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