title: Peptide-templated acyl transfer: a chemical method for the labeling of membrane proteins on live cells.
abstract: The development of a method is described for the chemical labeling of proteins which occurs with high target specificity, proceeds within seconds to minutes, and offers a free choice of the reporter group. The method relies upon the use of peptide templates, which align a thioester and an N-terminal cysteinyl residue such that an acyl transfer reaction is facilitated at nanomolar concentrations. The protein of interest is N-terminally tagged with a 22 aa long Cys-E3 peptide (acceptor), which is capable of forming a coiled-coil with a reporter-armed K3 peptide (donor). This triggers the transfer of the reporter to the acceptor on the target protein. Because ligation of the two interacting peptides is avoided, the mass increase at the protein of interest is minimal. The method is exemplified by the rapid fluorescent labeling and fluorescence microscopic imaging of the human Y2 receptor on living cells.
authors: Institut für Chemie, Humboldt-Universität zu Berlin, Brook-Taylor-Strasse 2, 12489 Berlin (Germany).
authors: Ulrike
authors: U
authors: Reinhardt
authors: Jonathan
authors: J
authors: Lotze
authors: Sarah
authors: S
authors: Zernia
authors: Karin
authors: K
authors: Mörl
authors: Annette G
authors: AG
authors: Beck-Sickinger
authors: Oliver
authors: O
authors: Seitz
mesh_terms: D002470:Cell Survival; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D013194:Staining and Labeling
chemical_list: D008565:Membrane Proteins; D010455:Peptides
keywords: bioorganic chemistry; fluorescent probes; membrane proteins; peptides; protein modifications