A genetically encoded multifunctional unnatural amino acid for versatile protein manipulations in living cells. Issue 12 (15th August 2016)

• Record Type: Journal Article
• Authors: Ge, Yun; Fan, Xinyuan; Chen, Peng R.
• Is Part Of: Chemical science. Volume 7:Issue 12(2016:Dec.)
• Page Start: 7055
• Page End: 7060
• Publication Date: 2016-08-15
• Subjects: Chemistry -- Periodicals; 540.5
• DOI: 10.1039/c6sc02615j
• Languages: English
• ISSNs: 2041-6520

Abstract

Abstract : A multifunctional UAA, PABK, is developed for diverse protein manipulation purposes, especially protein activation in living cells. Abstract : The genetic code expansion strategy allowed incorporation of unnatural amino acids (UAAs) bearing diverse functional groups into proteins, providing a powerful toolkit for protein manipulation in living cells. We report a multifunctional UAA, N ε - p -azidobenzyloxycarbonyl lysine (PABK), that possesses a panel of unique properties capable of fulfilling various protein manipulation purposes. In addition to being used as a bioorthogonal ligation handle, an infrared probe and a photo-affinity reagent, PABK was shown to be chemically decaged by trans -cyclooctenols via a strain-promoted 1, 3-dipolar cycloaddition, which provides a new bioorthogonal cleavage strategy for intracellular protein activation. The biocompatibility and efficiency of this method were demonstrated by decaging of a PABK-caged firefly luciferase under living conditions. We further extended this method to chemically rescue a bacterial toxin OspF inside mammalian host cells.