RS/tRNA Foundational Publication Support
Miyake-Stoner, Shigeki J, Andrew M Miller, Jared T Hammill, Jennifer C Peeler, Kenneth R Hess, Ryan A Mehl, and Scott H Brewer. (2009) 2009. “Probing Protein Folding Using Site-Specifically Encoded Unnatural Amino Acids As Fret Donors With Tryptophan.”. Biochemistry 48 (25): 5953-62. doi:10.1021/bi900426d.
Beyer, Jenna N, Parisa Hosseinzadeh, Ilana Gottfried-Lee, Elise M Van Fossen, Phillip Zhu, Riley M Bednar, Andrew Karplus, Ryan A Mehl, and Richard B Cooley. (2020) 2020. “Overcoming Near-Cognate Suppression In A Release Factor 1-Deficient Host With An Improved Nitro-Tyrosine Trna Synthetase.”. Journal Of Molecular Biology 432 (16): 4690-4704. doi:10.1016/j.jmb.2020.06.014.
RS/tRNA Pair Development Year
2009
ncAA(s) Incorporated
p-Cyano-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
FRET Probe, Infrared probe
p-Ethynyl-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
FRET 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.
RS Organism of Origin
Parent RS
RS Mutations
Y32L
L65V
F108W
Q109M
D158G
I159P
L65V
F108W
Q109M
D158G
I159P
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Other tRNA Mutations
C17A
U17aG
U20C
G37A
U47G
U17aG
U20C
G37A
U47G
Multiple tRNAs?
N/a
RS/tRNA Availability
On pDule plasmid as Addgene #85494
On pDule2 plasmid as Addgene #85495
O pEVOL plasmid as Addgene #164579 designed for use in dual encoding applications involving pAzido-Phe
On pDule2 plasmid as Addgene #85495
O pEVOL plasmid as Addgene #164579 designed for use in dual encoding applications involving pAzido-Phe
Used in what cell line?
RS/tRNA Additional Notes
Selected against para-cyano-Phe (pCNF) and encorporates pENPhe. With 1 mM L-pCNF generated T4 lysozyme(153) at ~7.5 mg/L (~25% effciency). With 1 mM racemic pENPhe, yielded 23 mg/L protein (~75% efficiency). Full fidelity was confirmed by MS.
Shown in 2019 foundational paper to produce full-fidelity incorporation of pAzF into sfGFP(150) in truncation free B95 RF1- E coli.
Shown in 2019 foundational paper to produce full-fidelity incorporation of pAzF into sfGFP(150) in truncation free B95 RF1- E coli.