RS/tRNA Foundational Publication Support
Ai, Hui-Wang, Weijun Shen, Amit Sagi, Peng R Chen, and Peter G Schultz. (2011) 2011. “Probing Protein-Protein Interactions With A Genetically Encoded Photo-Crosslinking Amino Acid.”. Chembiochem : A European Journal Of Chemical Biology 12 (12): 1854-7. doi:10.1002/cbic.201100194.
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 Usage Publications
Kleiner, Ralph E, Lisa E Hang, Kelly R Molloy, Brian T Chait, and Tarun M Kapoor. (2018) 2018. “A Chemical Proteomics Approach To Reveal Direct Protein-Protein Interactions In Living Cells.”. Cell Chemical Biology 25 (1): 110-120.e3. doi:10.1016/j.chembiol.2017.10.001.
Bartoschek, Michael D, Enes Ugur, Tuan-Anh Nguyen, Geraldine Rodschinka, Michael Wierer, Kathrin Lang, and Sebastian Bultmann. (2021) 2021. “Identification Of Permissive Amber Suppression Sites For Efficient Non-Canonical Amino Acid Incorporation In Mammalian Cells.”. Nucleic Acids Research 49 (11): e62. doi:10.1093/nar/gkab132.
RS/tRNA Pair Development Year
2011
ncAA(s) Incorporated
N6-[(2-(3-Methyl-3H-diazirin-3-yl)ethoxy)carbonyl]-L-lysine (AbK; DiAzKs)
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
Introduced as a photocrosslinking agent that was aliphatic rather than aromatic giving it more flexibility. Similar in structure to DiZPK.
RS Organism of Origin
Parent RS
RS Mutations
L274M
C313A
Y349F
C313A
Y349F
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Other tRNA Mutations
n/a
Multiple tRNAs?
The second foundational paper reported using this RS, but paired with a tRNA having an additional U25G mutation, which as reported in doi: 10.1021/bi4000244 improved its amber suppression efficiency.
RS/tRNA Availability
Available from Addgene #49086 (pDULE backbone from foundatinal paper 1) and #126035 (pEVOL backbone from foundational paper 2)
RS/tRNA Additional Notes
In E.coli DH10B, using 1 mM AbK yielded Myoglobin(99) at ~12 mg/L, confirmed by ESI-MS; no protein was detected in the absence of AbK. Photolysis upon exposure to 360 nm light for 10 min was also confirmed by MS. Again using 1mM AbK, HEK293T cells (with a pCMV-Abk RS plasmid) produced pEGFP(39) yielding ~0.4 mg from 1x10^7 cells with incorporation confirmed by ESI-MS.