RS/tRNA Foundational Publication Support
Meineke, Birthe, Johannes Heimgärtner, Lorenzo Lafranchi, and Simon J Elsässer. (2018) 2018. “Methanomethylophilus Alvus Mx1201 Provides Basis For Mutual Orthogonal Pyrrolysyl Trna/Aminoacyl-Trna Synthetase Pairs In Mammalian Cells.”. Acs Chemical Biology 13 (11): 3087-3096. doi:10.1021/acschembio.8b00571.
RS/tRNA Usage Publications
Meineke, Birthe, Johannes Heimgärtner, Jürgen Eirich, Michael Landreh, and Simon J Elsässer. (2020) 2020. “Site-Specific Incorporation Of Two Ncaas For Two-Color Bioorthogonal Labeling And Crosslinking Of Proteins On Live Mammalian Cells.”. Cell Reports 31 (12): 107811. doi:10.1016/j.celrep.2020.107811.
RS/tRNA Pair Development Year
2018
ncAA(s) Incorporated
4'-TCOK (TCO)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Reactive handle for IEDDA.
Nε-acetyl-L-lysine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
They are used for sites specifically studying post-translational modification acetylation events on particular lysine residues.
RS Organism of Origin
Parent RS
RS Mutations
Y126A
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Other tRNA Mutations
C41CA
RS/tRNA Availability
https://www.addgene.org/140023/
Used in what cell line?
RS/tRNA Additional Notes
Effective incorporation of TCO*K in dual-encoding alongside AcK which was done using a different RS/tRNA and plasmid. The combined yield in dual-encoding of GFP was determined to be ~1.5 μg per 25 mL/175m2 HEK293T culture. Incorporation was confirmed with ESI-MS, however tetrazine pull-down revealed that some amount of GFP did not contain TCO*K. Incorporation of TCO*K was done at site 102, Ack was incorporated at site 150.
Interestingly, the Mx1201 PylRS has evolved efficient tRNA binding through its catalytic domain, making an N-terminal domain obsolete.
Interestingly, the Mx1201 PylRS has evolved efficient tRNA binding through its catalytic domain, making an N-terminal domain obsolete.