RS/tRNA Foundational Publication Support
Cooley, Richard B, and Holger Sondermann. (2017) 2017. “Probing Protein-Protein Interactions With Genetically Encoded Photoactivatable Cross-Linkers.”. Methods In Molecular Biology (Clifton, N.j.) 1657: 331-345. doi:10.1007/978-1-4939-7240-1_26.
RS/tRNA Usage Publications
Pan, Dongqing, Kai Walstein, Annika Take, David Bier, Nadine Kaiser, and Andrea Musacchio. (2019) 2019. “Mechanism Of Centromere Recruitment Of The Cenp-A Chaperone Hjurp And Its Implications For Centromere Licensing.”. Nature Communications 10 (1): 4046. doi:10.1038/s41467-019-12019-6.
RS/tRNA Pair Development Year
2011
ncAA(s) Incorporated
N6-[(2-(3-Methyl-3H-diazirin-3-yl)ethoxy)carbonyl]-L-lysine
ncAA Structure (png, jpg, jpeg)
![N6-[(2-(3-Methyl-3H-diazirin-3-yl)ethoxy)carbonyl]-L-lysine](/sites/gce4all/files/styles/medium/public/2025-01/Page_20_Structure_6.png?itok=YosSmxRs)
ncAA Utility
Photo-cross-linker
RS Organism of Origin
Parent RS
RS Mutations
Leu274Met
Cys313Ala
Tyr349Phe
Cys313Ala
Tyr349Phe
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Other tRNA Mutations
n/a
Multiple tRNAs?
n/a
RS/tRNA Availability
Available from Addgene #49086
RS/tRNA Additional Notes
Tested in E.coli DH10B cell line by incorporating site specifically at residue 99 of full-length Myoglobin. An yield of 12±1.1 mg per liter of culture. Protein presence was observed using ESI-MS; no protien presence was dected in the absence of 1mM Abk. Photolysis activity was confirmed using mass spectrometry after exposure to 360 nm light for 10 min.
Testing in HEK293T cell line was performed using pCMV-Abk RS, while pEGFP-Tyr39TAG was used as the florescent reporter. EGFP imaged using flourscence microsopy after expression in presence or absence of 1mM Abk. The reporter was only observed in the presence of Abk. An yield of ~0.4mg from 1x10^7 cell was reported, after being subjected to 360nm light for 10 min with the incorporation being confirmed using ESI-MS
Testing in HEK293T cell line was performed using pCMV-Abk RS, while pEGFP-Tyr39TAG was used as the florescent reporter. EGFP imaged using flourscence microsopy after expression in presence or absence of 1mM Abk. The reporter was only observed in the presence of Abk. An yield of ~0.4mg from 1x10^7 cell was reported, after being subjected to 360nm light for 10 min with the incorporation being confirmed using ESI-MS