title: Nanomolar affinity protein trans-splicing monitored in real-time by fluorophore-quencher pairs.
abstract: High background originating from non-reacted, 'always-on' fluorescent probes remains a key unsolved problem in life science since washing procedures are not easily applicable. Covalent labeling approaches with simultaneous activation of fluorescence are advantageous to increase sensitivity and to reduce background signal. Here, we combined high-affinity protein trans-splicing with fluorophore/quencher pairs for online detection of covalent N-terminal 'traceless' protein labeling under physiological conditions in cellular environments. Substantial fluorescence enhancement at nanomolar probe concentrations was achieved.
authors.affiliation: Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt a.M., Germany. tampe@em.uni-frankfurt.de.
authors.forename: M
authors.initials: M
authors.lastname: Braner
authors.affiliation: Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt a.M., Germany. tampe@em.uni-frankfurt.de.
authors.forename: R
authors.initials: R
authors.lastname: Wieneke
authors.affiliation: Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue-Str. 9, 60438 Frankfurt a.M., Germany. tampe@em.uni-frankfurt.de.
authors.forename: R
authors.identifier: 0000-0002-0403-2160
authors.initials: R
authors.lastname: Tampé
mesh_terms: D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D013997:Time Factors; D020040:Trans-Splicing
chemical_list: D005456:Fluorescent Dyes; D011506:Proteins