title: Semisynthesis of fluorescent metabolite sensors on cell surfaces.
abstract: Progress in understanding signal transduction and metabolic pathways is hampered by a shortage of suitable sensors for tracking metabolites, second messengers, and neurotransmitters in living cells. Here we introduce a class of rationally designed semisynthetic fluorescent sensor proteins, called Snifits, for measuring metabolite concentrations on the cell surface of mammalian cells. Functional Snifits are assembled on living cells through two selective chemical labeling reactions of a genetically encoded protein scaffold. Our best Snifit displayed fluorescence intensity ratio changes on living cells significantly higher than any previously reported cell-surface-targeted fluorescent sensor protein. This work establishes a generally applicable and rational strategy for the generation of cell-surface-targeted fluorescent sensor proteins for metabolites of interest.
authors: Institute of Chemical Sciences and Engineering, National Centre of Competence in Research Chemical Biology, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
authors: Matthias A
authors: MA
authors: Brun
authors: Rudolf
authors: R
authors: Griss
authors: Luc
authors: L
authors: Reymond
authors: Kui-Thong
authors: KT
authors: Tan
authors: Joachim
authors: J
authors: Piguet
authors: Ruud J R W
authors: RJ
authors: Peters
authors: Horst
authors: H
authors: Vogel
authors: Kai
authors: K
authors: Johnsson
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D008164:Luminescent Proteins