scholarly journals Genetic Encoding of Photocaged Cysteine Allows Photoactivation of TEV Protease in Live Mammalian Cells

2014 ◽  
Vol 136 (6) ◽  
pp. 2240-2243 ◽  
Author(s):  
Duy P. Nguyen ◽  
Mohan Mahesh ◽  
Simon J. Elsässer ◽  
Susan M. Hancock ◽  
Chayasith Uttamapinant ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Tev Protease ◽  
Genetic Encoding

Photoactivation of Innate Immunity Receptor TLR8 in Live Mammalian Cells by Genetic Encoding of Photocaged Tyrosine

ChemBioChem ◽  
2021 ◽  
Author(s):  
Yi Yang ◽  
Shuchen Luo ◽  
Jian Huang ◽  
Yu Xiao ◽  
Yixuan Fu ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Mammalian Cells ◽  
Genetic Encoding

scholarly journals Functional genetic encoding of sulfotyrosine in mammalian cells

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Xinyuan He ◽  
Yan Chen ◽  
Daisy Guiza Beltran ◽  
Maia Kelly ◽  
Bin Ma ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Biochemical Characterization ◽  
Trna Synthetase ◽  
Functional Verification ◽  
Cellular Processes ◽  
Genetic Encoding ◽  
Efficient Expression

Abstract Protein tyrosine O-sulfation (PTS) plays a crucial role in extracellular biomolecular interactions that dictate various cellular processes. It also involves in the development of many human diseases. Regardless of recent progress, our current understanding of PTS is still in its infancy. To promote and facilitate relevant studies, a generally applicable method is needed to enable efficient expression of sulfoproteins with defined sulfation sites in live mammalian cells. Here we report the engineering, in vitro biochemical characterization, structural study, and in vivo functional verification of a tyrosyl-tRNA synthetase mutant for the genetic encoding of sulfotyrosine in mammalian cells. We further apply this chemical biology tool to cell-based studies on the role of a sulfation site in the activation of chemokine receptor CXCR4 by its ligand. Our work will not only facilitate cellular studies of PTS, but also paves the way for economical production of sulfated proteins as therapeutic agents in mammalian systems.

Genetic Encoding of Caged Cysteine and Caged Homocysteine in Bacterial and Mammalian Cells

ChemBioChem ◽  
2014 ◽  
Vol 15 (12) ◽  
pp. 1793-1799 ◽  
Author(s):  
Rajendra Uprety ◽  
Ji Luo ◽  
Jihe Liu ◽  
Yuta Naro ◽  
Subhas Samanta ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Genetic Encoding

scholarly journals Engineered tobacco etch virus (TEV) protease active in the secretory pathway of mammalian cells

2015 ◽  
Vol 212 ◽  
pp. 159-166 ◽  
Author(s):  
Francesca Cesaratto ◽  
Alejandro López-Requena ◽  
Oscar R. Burrone ◽  
Gianluca Petris
Keyword(s):  
Mammalian Cells ◽  
Secretory Pathway ◽  
Tobacco Etch Virus ◽  
Tev Protease

scholarly journals Genetic encoding of a highly photostable, long lifetime fluorescent amino acid for imaging in mammalian cells

2021 ◽  
Author(s):  
Chloe M. Jones ◽  
D. Miklos Robkis ◽  
Robert J. Blizzard ◽  
Mika Munari ◽  
Yarra Venkatesh ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Mammalian Cells ◽  
Trna Synthetase ◽  
Fluorescence Lifetime Imaging ◽  
High Quantum Yield ◽  
Lifetime Imaging ◽  
Genetic Encoding ◽  
Long Lifetime ◽  
Fluorescent Amino Acid ◽  
Genetic Incorporation

Acridonylalanine (Acd) is photostable, with a high quantum yield and long fluorescence lifetime in water. An evolved tRNA synthetase (RS) enables genetic incorporation of Acd in mammalian cells and its use in fluorescence lifetime imaging microscopy.

Photocontrol of Tyrosine Phosphorylation in Mammalian Cells via Genetic Encoding of Photocaged Tyrosine

2012 ◽  
Vol 134 (29) ◽  
pp. 11912-11915 ◽  
Author(s):  
Eyal Arbely ◽  
Jessica Torres-Kolbus ◽  
Alexander Deiters ◽  
Jason W. Chin
Keyword(s):  
Tyrosine Phosphorylation ◽  
Mammalian Cells ◽  
Genetic Encoding

scholarly journals Front Cover: Reversible and Tunable Photoswitching of Protein Function through Genetic Encoding of Azobenzene Amino Acids in Mammalian Cells (ChemBioChem 20/2018)

ChemBioChem ◽  
2018 ◽  
Vol 19 (20) ◽  
pp. 2120-2120
Author(s):  
Ji Luo ◽  
Subhas Samanta ◽  
Marino Convertino ◽  
Nikolay V. Dokholyan ◽  
Alexander Deiters
Keyword(s):  
Amino Acids ◽  
Protein Function ◽  
Mammalian Cells ◽  
Front Cover ◽  
Genetic Encoding

scholarly journals Time-gated detection of protein-protein interactions with transcriptional readout

2017 ◽  
Author(s):  
Min Woo Kim ◽  
Wenjing Wang ◽  
Mateo I. Sanchez ◽  
Robert Coukos ◽  
Mark Von Zastrow ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Mammalian Cells ◽  
Time Window ◽  
High Specificity ◽  
Large Cell ◽  
Protein Protein Interactions ◽  
Signal Ratio ◽  
High Background ◽  
Tev Protease ◽  
Fast Light

AbstractTranscriptional assays such as yeast two hybrid, split ubiquitin, and Tango that convert transient protein-protein interactions (PPIs) in cells into stable expression of transgenes are powerful tools for PPI discovery, high-throughput screens, and analysis of large cell populations. However, these assays frequently suffer from high background and they lose all information about PPI dynamics. To address these limitations, we developed a light-gated transcriptional assay for PPI detection called PPI-FLARE (PPI-Fast Light- and Activity-Regulated Expression). PPI-FLARE requires both a PPI to deliver TEV protease proximal to its cleavage peptide, and externally-applied blue light to uncage the cleavage peptide, in order to release a membrane-tethered transcription factor (TF) for translocation to the nucleus. We used PPI-FLARE to detect the ligand-induced association of 12 different PPIs in living mammalian cells, with a temporal resolution of 5 minutes and a ±ligand signal ratio up to 37. By systematically shifting the light irradiation window, we could reconstruct PPI time-courses, distinguishing between GPCRs that engage in transient versus sustained interactions with the cytosolic effector arrestin. When combined with FACS, PPI-FLARE enabled >100-fold enrichment of cells experiencing a specific GPCR-arrestin PPI during a short 10-minute light window over cells missing that PPI during the same time window. Due to its high specificity, sensitivity, and generality, PPI-FLARE should be a broadly useful tool for PPI analysis and discovery.

Sign in / Sign up

Export Citation Format

Share Document