RS/tRNA Foundational Publication Support
Hao, Ziyang, Yanqun Song, Shixian Lin, Maiyun Yang, Yujie Liang, Jing Wang, and Peng R Chen. (2011) 2011. “A Readily Synthesized Cyclic Pyrrolysine Analogue For Site-Specific Protein "Click" Labeling.”. Chemical Communications (Cambridge, England) 47 (15): 4502-4. doi:10.1039/c1cc00024a.
Li, Yiming, Maiyun Yang, Yichao Huang, Xiaoda Song, Lei Liu, and Peng R. Chen. (0BC) 2012. “Genetically Encoded Alkenyl–Pyrrolysine Analogues For Thiol–Ene Reaction Mediated Site-Specific Protein Labeling”. Chemical Science 3 (9): 2766-2770, . doi:10.1039/C2SC20433A.
Li, Yiming, Man Pan, Yitong Li, Yichao Huang, and Qingxiang Guo. (2013) 2013. “Thiol-Yne Radical Reaction Mediated Site-Specific Protein Labeling Via Genetic Incorporation Of An Alkynyl-L-Lysine Analogue.”. Organic & Biomolecular Chemistry 11 (16): 2624-9. doi:10.1039/c3ob27116a.
RS/tRNA Usage Publications
Li, Yiming, Man Pan, Yitong Li, Yichao Huang, and Qingxiang Guo. (2013) 2013. “Thiol-Yne Radical Reaction Mediated Site-Specific Protein Labeling Via Genetic Incorporation Of An Alkynyl-L-Lysine Analogue.”. Organic & Biomolecular Chemistry 11 (16): 2624-9. doi:10.1039/c3ob27116a.
RS/tRNA Pair Development Year
2011
ncAA(s) Incorporated
N6-((2-azidoethoxy)carbonyl)-L-lysine(AzeoK)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Reactive handle for azide-alkyne cycloaddition
N6-(((1R,2R)-2-azidocyclopentyloxy)carbonyl)-L-lysine (ACPK)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
contains reactive azido group
BlocLys
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Reactive handle for thiol-ene reaction
PlocLys
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Reactive handle for thiol-ene reaction
N6-((Pent-4-yn-1-yloxy)carbonyl)-L-lysine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Reactive handle for thiol-yne reaction
RS Organism of Origin
Parent RS
RS Mutations
L274A
C313V
Y349F
C313V
Y349F
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
RS/tRNA Availability
N/A
RS/tRNA Additional Notes
In E.coli BL21-DE3 cells, produced GFP(149) ACPK at ~10mg/L, with fidelity confirmed by MS. Also incorporated into HdeA(58) and showed CuAAC could be used for click labeling in live bacteria. Incorporation in HEK293 cells successfully produced a p53EGFP(372) protein with fluorescence only observed when cells were supplemented by 1mM ACPK. Expression efficiency was ~50% of WT levels. A couple additional applications were shown. Also, a second evolved RS (ACPK-RSa) with the same mutations except but C313A rather than V showed lower efficiency in E coli.
In second foundational paper, this RS was shown to even more efficiently incorporate alkene containing lys derivatives "BLOC" and "PLOC" with incorporation confirmed by MS, and the incorporated ncAAs shown to be avaialble for thiol-ene ligation reactions. An "ALOC" Lys derivative 1-carbon shorter than BLOC was also incorporated but less well.
Third foundational paper reports highly efficient incorporation of "pent-4-yne" Lys derivative useful for radical thiol-yne dual labeling. Also incorporated were alkyne-Lys derivatives one and two methylene groups shorter.
In second foundational paper, this RS was shown to even more efficiently incorporate alkene containing lys derivatives "BLOC" and "PLOC" with incorporation confirmed by MS, and the incorporated ncAAs shown to be avaialble for thiol-ene ligation reactions. An "ALOC" Lys derivative 1-carbon shorter than BLOC was also incorporated but less well.
Third foundational paper reports highly efficient incorporation of "pent-4-yne" Lys derivative useful for radical thiol-yne dual labeling. Also incorporated were alkyne-Lys derivatives one and two methylene groups shorter.