Genetically encoded fluorescent unnatural amino acids and FRET probes for detecting deubiquitinase activities
Manjia Li, Feifei Wang, Long Yan, Minghao Lu, Yuqing Zhang, Tao Peng
Abstract
Herein we present the genetic encoding of 7-aminocoumarin-based lysine derivatives, ACouK and AFCouK, into proteins in both bacterial and mammalian cells and the characterization of FRET pairs comprising ACouK or AFCouK as the donor and GFP as the acceptor. We further report the application of the FRET pairs to construct fully genetically encoded ratiometric probes for detecting deubiquitinases and screening for inhibitors.
Topics & Concepts
Förster resonance energy transferChemistryFluorescenceGreen fluorescent proteinLysineDeubiquitinating enzymeAmino acidFluorescent proteinBiochemistryComputational biologyAcceptorCombinatorial chemistryGeneBiologyUbiquitinQuantum mechanicsCondensed matter physicsPhysicsUbiquitin and proteasome pathwaysClick Chemistry and ApplicationsGlycosylation and Glycoproteins Research