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Peptide‐Templated Acyl Transfer: A Chemical Method for the Labeling of Membrane Proteins on Live Cells
Author(s) -
Reinhardt Ulrike,
Lotze Jonathan,
Zernia Sarah,
Mörl Karin,
BeckSickinger Annette G.,
Seitz Oliver
Publication year - 2014
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201403214
Subject(s) - peptide , native chemical ligation , chemistry , thioester , residue (chemistry) , fluorescence , green fluorescent protein , coiled coil , biochemistry , membrane , biophysics , combinatorial chemistry , cysteine , biology , enzyme , gene , physics , quantum mechanics
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 Y 2 receptor on living cells.