Torres-Kolbus, J and Chou, C and Liu, J and Deiters, A
(2014)
Synthesis of non-linear protein dimers through a genetically encoded thiol-ene reaction.
PLoS ONE, 9 (9).
Abstract
Site-specific incorporation of bioorthogonal unnatural amino acids into proteins provides a useful tool for the installation of specific functionalities that will allow for the labeling of proteins with virtually any probe. We demonstrate the genetic encoding of a set of alkene lysines using the orthogonal PylRS/PylTCUA pair in Escherichia coli. The installed double bond functionality was then applied in a photoinitiated thiol-ene reaction of the protein with a fluorescent thiol-bearing probe, as well as a cysteine residue of a second protein, showing the applicability of this approach in the formation of heterogeneous non-linear fused proteins. © 2014 Torres-Kolbus et al.
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