Rankings
Publications
Sources
Publishers
Scholars
Organizations
About
Login
Register
Home
Publications
Faculty Opinions recommendation of Proximity-triggered covalent stabilization of low-affinity protein complexes in vitro and in vivo.
Home
Publications
Faculty Opinions recommendation of Proximity-triggered covalent stabilization of low-affinity protein complexes in vitro and in vivo.
Faculty Opinions recommendation of Proximity-triggered covalent stabilization of low-affinity protein complexes in vitro and in vivo.
GO
Gottfried Otting
Gottfried Otting
Publisher Website
Google Scholar
Cite
Download
Share
Download
6 October 2017
dataset
Published by
H1 Connect
Vol. 56
(49)
https://doi.org/10.3410/f.731430647.793537477
Abstract
The characterization of low-affinity protein complexes is challenging due to their dynamic nature. Here we present a method to stabilize transient protein complexes in vivo by generating a covalent and conformationally flexible bridge between the interaction partners. A highly active pyrrolysyl tRNA synthetase mutant directs the incorporation of unnatural amino acids bearing bromoalkyl moieties (BrCnK) into proteins. We demonstrate for the first time that low-affinity protein complexes between BrCnK-containing proteins and their binding partners can be stabilized in vivo in bacterial and mammalian cells. Using this approach we determined the crystal structure of a transient GDP-bound complex between a small G-protein and its nucleotide exchange factor. We envision that this approach will prove valuable as a general tool for validating and characterizing protein-protein interactions in vitro and in vivo. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Cited by 1 article