RS/tRNA Foundational Publication Support
Bryson, David I., Chenguang Fan, Li-Tao Guo, Corwin Miller, Dieter Söll, and David R. Liu. (dec) 2017. “Continuous Directed Evolution Of Aminoacyl-Trna Synthetases”. Nature Chemical Biology 13: 1253-1260. doi:10.1038/nchembio.2474.
RS/tRNA Usage Publications
Stieglitz, Jessica T., Priyanka Lahiri, Matthew I. Stout, and James A. Van Deventer. (may) 2022. “Exploration Of Methanomethylophilus Alvus Pyrrolysyl-Trna Synthetase Activity In Yeast”. Acs Synthetic Biology 11: 1824-1834. doi:10.1021/acssynbio.2c00001.
RS/tRNA Pair Development Year
2017
ncAA(s) Incorporated
Nε-acetyl-L-lysine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
They are used for sites specifically studying post-translational modification acetylation events on particular lysine residues.
RS Organism of Origin
Parent RS
RS Mutations
V31I
T56P
H62Y
D76G
A100E
L266M
L270I
Y271F
L274A
C313F
T56P
H62Y
D76G
A100E
L266M
L270I
Y271F
L274A
C313F
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Other tRNA Mutations
N/A
Multiple tRNAs?
unclear from foundational paper if this work was done with the Pyl tRNA from Mm or Mb
RS/tRNA Availability
AddGene Plasmid #104069
RS/tRNA Additional Notes
Created to test if adding the "IPYE" mutations to other PylRSs would improve their activity. This RS is a chimera of residues 1–149 of MbPylRS and 185–454 of MmPylRS, and the mutations taken from the MbAcKRS3 synthetase listed here as L266M, L270I, Y271F, L274A and C313F are based on the Mb residue numbering; the mutations based on their Mm residue numbers are L301M, L305I, Y306F, L309A and C348F.
In the presence of 1 mM AcK, adding the IPYE mutations gave an ~8-fold increased efficiency of expression of a reporter sfGFP(2) (reaching ~22% efficiency) compared to that achieved by chAcK3RS (i.e. without the IPYE mutations). This efficiency was higher than that achieved by the MbAcK3RS(IPYE) (~16%) or the MmAcK3RS(IPYE) (~12%).
In the presence of 1 mM AcK, adding the IPYE mutations gave an ~8-fold increased efficiency of expression of a reporter sfGFP(2) (reaching ~22% efficiency) compared to that achieved by chAcK3RS (i.e. without the IPYE mutations). This efficiency was higher than that achieved by the MbAcK3RS(IPYE) (~16%) or the MmAcK3RS(IPYE) (~12%).