RS/tRNA Foundational Publication Support
Zhang, Meng, Shixian Lin, Xinwen Song, Jun Liu, Ye Fu, Xi Ge, Xinmiao Fu, Zengyi Chang, and Peng R Chen. (2011) 2011. “A Genetically Incorporated Crosslinker Reveals Chaperone Cooperation In Acid Resistance.”. Nature Chemical Biology 7 (10): 671-7. doi:10.1038/nchembio.644.
RS/tRNA Pair Development Year
2011
ncAA(s) Incorporated
(3-(3-Methyl-3H diazirine-3-yl)-propaminocarbonyl- Nε-L-lysine ( DiZPK)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Photo-crosslinker
RS Organism of Origin
Parent RS
RS Mutations
L274A
C313S
C313S
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Other tRNA Mutations
n/a
RS/tRNA Availability
RS available in AddGene Plasmid #91705 (in a pSupAR vector backbone with a Cm resistance gene)
and Plasmid #91706 (for mammalian cells with a pCMV vector backbone)
and Plasmid #91706 (for mammalian cells with a pCMV vector backbone)
RS/tRNA Additional Notes
Using 1 mM DiZPK ncAA, produces myoglobin(99) at ~9 mg/L with fidelity confirmed by ESI-MS. In CHO cells, produces full-length EGFP(37). A second selected RS (DZKRS-a differing only in that it had a C313A rather than C313S mutation) worked but had somewhat lower efficiency of ncAA incorporation. The ncAA was incorporated into the E coli HdeA protein at multiple sites, and in those contexts shown to be a higher efficiency crosslinker than Bpa (~80% vs ~40% in one of the cases).