4654-14-2Relevant academic research and scientific papers
Methods and compositions for protein labeling using lipoic acid ligases
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, (2016/04/05)
The present disclosure provides compositions and methods of use thereof for labeling peptide and proteins in vitro or in vivo. The methods described herein employ lipoic acid ligase or mutants thereof, and lipoic acid analogs (e.g., lipoic acid analogs comprising a resorufin moiety) recognized by lipoic acid ligase and lipoic acid ligase mutants. Also provided herein is a method of imaging protein-protein interaction via a reaction mediated by lipoic acid ligase.
Computational design of a red fluorophore ligase for site-specific protein labeling in living cells
Liu, Daniel S.,Nivón, Lucas G.,Richter, Florian,Goldman, Peter J.,Deerinck, Thomas J.,Yao, Jennifer Z.,Richardson, Douglas,Phipps, William S.,Ye, Anne Z.,Ellisman, Mark H.,Drennan, Catherine L.,Baker, David,Ting, Alice Y.
, p. E4551 - E4559 (2015/02/19)
Chemical fluorophores offer tremendous size and photophysical advantages over fluorescent proteins but are much more challenging to target to specific cellular proteins. Here, we used Rosetta-based computation to design a fluorophore ligase that accepts the red dye resorufin, starting from Escherichia coli lipoic acid ligase. X-ray crystallography showed that the design closely matched the experimental structure. Resorufin ligase catalyzed the site-specific and covalent attachment of resorufin to various cellular proteins genetically fused to a 13-aa recognition peptide in multiple mammalian cell lines and in primary cultured neurons. We used resorufin ligase to perform superresolution imaging of the intermediate filament protein vimentin by stimulated emission depletion and electron microscopies. This work illustrates the power of Rosetta for major redesign of enzyme specificity and introduces a tool for minimally invasive, highly specific imaging of cellular proteins by both conventional and superresolution microscopies.
C-H amination in synthesis: An approach to the assembly of the B/C/D ring system of aconitine
Conrad, Rosemary M.,Du Bois
, p. 5465 - 5468 (2008/09/17)
A strategy for the preparation of aconitine is described that attempts to exploit chemoselective C-H amination and the electrophilic reactivity of oxathiazinane N,O-acetals for assembling the complex, polycyclic carbon framework of the natural product.
