RS/tRNA Foundational Publication Support
Coin, Irene, Vsevolod Katritch, Tingting Sun, Zheng Xiang, Fai Siu, Michael Beyermann, Raymond C Stevens, and Lei Wang. (2013) 2013. “Genetically Encoded Chemical Probes In Cells Reveal The Binding Path Of Urocortin-I To Crf Class B Gpcr.”. Cell 155 (6): 1258-69. doi:10.1016/j.cell.2013.11.008.
Takimoto, Jeffrey K, Katrina Adams, Zheng Xiang, and Lei Wang. (2009) 2009. “Improving Orthogonal Trna-Synthetase Recognition For Efficient Unnatural Amino Acid Incorporation And Application In Mammalian Cells.”. Molecular Biosystems 5 (9): 931-4. doi:10.1039/b904228h.
RS/tRNA Usage Publications
Seidel, Lisa, Barbara Zarzycka, Saheem Zaidi, Vsevolod Katritch, and Irene Coin. (2017) 2017. “Structural Insight Into The Activation Of A Class B G-Protein-Coupled Receptor By Peptide Hormones In Live Human Cells.”. Elife 6. doi:10.7554/eLife.27711.
RS/tRNA Pair Development Year
2013
ncAA(s) Incorporated
p-azido-L-phenylalanine (pAzF)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Used as a photocrosslinker, allowing for crosslinking and bioorthogonal ligation of protein.
RS Organism of Origin
Parent RS
RS Mutations
Y37L
D182S
F183M
L186A
D265R
D182S
F183M
L186A
D265R
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Multiple tRNAs?
Original: E.coli Tyr-tRNA
Mutated: B. stearothermophylus Tyr tRNA
Mutated: B. stearothermophylus Tyr tRNA
RS/tRNA Availability
Can be ordered from AddGene Plasmid #105829
Used in what cell line?
RS/tRNA Additional Notes
Was developed to improve the efficiency of the E2AziRS in mammalian cells by introduction of Bst-Yam Tyr-tRNA. Particularly the AddGene plasmid contain 3 cassates of the tRNA.
By testing specific crosslinking of native Ucn1 by N123Azi-CRF1R revealed ~30% yeild of the WT, while no crosslinking was observed in the absence of ligand or WT CRF1R.
By testing specific crosslinking of native Ucn1 by N123Azi-CRF1R revealed ~30% yeild of the WT, while no crosslinking was observed in the absence of ligand or WT CRF1R.