RS/tRNA Foundational Publication Support
Schultz, Kathryn C, Lubica Supekova, Youngha Ryu, Jianming Xie, Roshan Perera, and Peter G Schultz. (2006) 2006. “A Genetically Encoded Infrared Probe.”. Journal Of The American Chemical Society 128 (43): 13984-5.
Young, Douglas D, Travis S Young, Michael Jahnz, Insha Ahmad, Glen Spraggon, and Peter G Schultz. (2011) 2011. “An Evolved Aminoacyl-Trna Synthetase With Atypical Polysubstrate Specificity.”. Biochemistry 50 (11): 1894-900. doi:10.1021/bi101929e.
Chatterjee, Abhishek, Sophie B Sun, Jennifer L Furman, Han Xiao, and Peter G Schultz. (2013) 2013. “A Versatile Platform For Single- And Multiple-Unnatural Amino Acid Mutagenesis In Escherichia Coli.”. Biochemistry 52 (10): 1828-37. doi:10.1021/bi4000244.
Wilkinson, Henry C, and Paul A Dalby. (2021) 2021. “Fine-Tuning The Activity And Stability Of An Evolved Enzyme Active-Site Through Noncanonical Amino-Acids.”. The Febs Journal 288 (6): 1935-1955. doi:10.1111/febs.15560.
RS/tRNA Pair Development Year
2006
ncAA(s) Incorporated
p-cyano-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
FRET Probe (with Trp) and infrared probe
p-azido-L-phenylalanine (pAzF)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Used as a photocrosslinker, allowing for crosslinking and bioorthogonal ligation of protein.
p-iodo-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Mutant GFPexcitation/ emission, later used for protein crystallization and as reactive handle. Structural determination of proteins. Amber suppression in a TB model mycobacterium
p-bromo-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Mutant GFP
excitation/ emission
excitation/ emission
p-acetyl-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Site-directed spin labeling, electron paramagnetic resonance
p-isopropyl-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
tyrosine analog
para-nitro-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
n/a
O-allyl-L-tyrosine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
in E. coli expressed sfGFP2TAG at 10 mg/L using GMML media
p-ethynyl-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
FRET Probe
BiphenylAla(BipAla)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Synthetase intermediate developed on way to BpyAla incorporation
O-tert-butyl-L-tyrosine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
n/a
O-propargyl-L-tyrosine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Reactive handle for azide-alkyne cycloaddition
p-chloro-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
NMR probe
p-fluoro-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
NMR probe
p-trifluoroacetyl-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Flourescent Probe, Bioorthogonal ligation handle
O-benzyl-L-serine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
n/a
RS Organism of Origin
Parent RS
RS Mutations
Y32L
L65V
F108W
D158G
I159A
L65V
F108W
D158G
I159A
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Other tRNA Mutations
C17A
U17aG
U20C
G37A
U47G
U17aG
U20C
G37A
U47G
RS/tRNA Availability
pULTRA-CNF plasmid with this RS is available as Addgene Plasmid #48215 (developed in 2013 foundational paper for use in dual encoding)
Used in what cell line?
RS/tRNA Additional Notes
In presence of 2 mM pCNF, MS confirmed full-fidelity incorporation into Z-domain(7). Also installed into myoglobin(64) yielding 30 mg/L, the pCNF provided a useful infrared probe of ligand binding.
2011 foundational paper showed this RS to be unusually permissive (i.e. polyspecific) and identified 18 ncAAs that were all incorporated into myoglobin(107) at between 43 and 96% efficiency (w/ WT expression being ~16 mg/L). Also reported a crystal structure with bound pCNF.
In 2013, putting into pUltra plasmid led to 1.3-2-fold efficiency gains for various ncAAs.
2021 paper incorporated p-nitro, p-cyano and p-aminoPhe into a transketolase in BL21 and C321.ΔA.exp cells with careful MS analyses indicating both cell lines yielded roughly 80%, 75% and 50% fidelity, respectively.
2011 foundational paper showed this RS to be unusually permissive (i.e. polyspecific) and identified 18 ncAAs that were all incorporated into myoglobin(107) at between 43 and 96% efficiency (w/ WT expression being ~16 mg/L). Also reported a crystal structure with bound pCNF.
In 2013, putting into pUltra plasmid led to 1.3-2-fold efficiency gains for various ncAAs.
2021 paper incorporated p-nitro, p-cyano and p-aminoPhe into a transketolase in BL21 and C321.ΔA.exp cells with careful MS analyses indicating both cell lines yielded roughly 80%, 75% and 50% fidelity, respectively.