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 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
L301M
L305I
Y306F
L309A
C348F
T56P
H62Y
D76G
A100E
L301M
L305I
Y306F
L309A
C348F
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Multiple tRNAs?
Not clear from the foundational paper or the Addgene listing if this was used with the Mb or the Mm tRNA
RS/tRNA Availability
Addgene plasmid #104071
RS/tRNA Additional Notes
Created to test if adding the "IPYE" mutations to various PylRSs would improve their activity. In this case, the IPYE mutations were added to an MmPylRS into which were also transplanted the mutations present in MbAcKRS3. The transplant without the IPYE led to what was called MmAcK3RS, and important to note is that it is a different enzyme from the MmAcKRSs that were selected from scratch (see MmAcKRS1 and MmAcKRS2).
For the active-site-transplant MmAcK3RS, the addition of the IPYE resulted in an ~10-fold increased efficiency of expression of a reporter sfGFP(2) (reaching ~12% efficiency). This efficiency was a lower than that achieved by the chAcK3RS(IPYE) (~22%) or the MbAcK3RS(IPYE) (~17%).
Note: unclear from the papers is if the D76G mutation originally present in MbAcKRS-1 and listed as a mutation is still present.
For the active-site-transplant MmAcK3RS, the addition of the IPYE resulted in an ~10-fold increased efficiency of expression of a reporter sfGFP(2) (reaching ~12% efficiency). This efficiency was a lower than that achieved by the chAcK3RS(IPYE) (~22%) or the MbAcK3RS(IPYE) (~17%).
Note: unclear from the papers is if the D76G mutation originally present in MbAcKRS-1 and listed as a mutation is still present.