biostudies-literatureUnknown12(14)Sun XThe ability to specifically attach chemical probes to individual proteins represents a powerful approach to the study and manipulation of protein function in living cells. It provides a simple, robust and versatile approach to the imaging of fusion proteins in a wide range of experimental settings. However, a potential drawback of detection using chemical probes is the fluorescence background from unreacted or nonspecifically bound probes. In this report we present the design and application of novel fluorogenic probes for labeling SNAP-tag fusion proteins in living cells. SNAP-tag is an engineered variant of the human repair protein O(6)-alkylguanine-DNA alkyltransferase (hAGT) that covalently reacts with benzylguanine derivatives. Reporter groups attached to the benzyl moiety become covaChembiochem : a European journal of chemical biology2217-26https://www.ebi.ac.uk/biostudies/studies/S-EPMC3213346biostudies-literatureDevelopment of SNAP-tag fluorogenic probes for wash-free fluorescence imaging.PMC3213346Sun LHoward AJohnsson KNoren CJBuswell JMasharina ACorrea IRXu MQSun XZhang ABaker BMaurel DfalseDevelopment of SNAP-tag fluorogenic probes for wash-free fluorescence imaging.The ability to specifically attach chemical probes to individual proteins represents a powerful approach to the study and manipulation of protein function in living cells. It provides a simple, robust and versatile approach to the imaging of fusion proteins in a wide range of experimental settings. However, a potential drawback of detection using chemical probes is the fluorescence background from unreacted or nonspecifically bound probes. In this report we present the design and application of novel fluorogenic probes for labeling SNAP-tag fusion proteins in living cells. SNAP-tag is an engineered variant of the human repair protein O(6)-alkylguanine-DNA alkyltransferase (hAGT) that covalently reacts with benzylguanine derivatives. Reporter groups attached to the benzyl moiety become cova2011-01-01T00:00:00Z2011 Sep2025-04-05T09:14:06.771Z2019-03-27T00:45:54ZS-EPMC32133462179315010.1002/cbic.201100173