Journal
CHEMBIOCHEM
Volume 18, Issue 14, Pages 1442-1447Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201700147
Keywords
genetic code expansion; protease; protein engineering; synthetic biology; unnatural amino acid
Funding
- National Science Foundation [CBET-1603930]
- Charles E. Kaufman Foundation of Pittsburgh Foundation
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [1603930] Funding Source: National Science Foundation
Ask authors/readers for more resources
We genetically encoded three new caged tyrosine analogues with improved photochemical properties by using an engineered pyrrolysyl-tRNA synthetase/tRNA(CUA) pair in bacterial and mammalian cells. We applied the new tyrosine analogues to the photoregulation of firefly luciferase by caging its key tyrosine residue, Tyr340, and observed excellent off-to-on light switching. This reporter was then used to evaluate the activation rates of the different light-removable protecting groups in live cells. We identified the nitropiperonyl caging group as an excellent compromise between incorporation efficiency and photoactivation properties. To demonstrate applicability of the new caged tyrosines, an important proteolytic enzyme, tobacco etch virus (TEV) protease, was engineered for optical control. The ability to incorporate differently caged tyrosine analogues into proteins in live cells further expands the unnatural amino acid and optogenetic toolbox.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available