RS/tRNA Foundational Publication Support
Chin, Jason W., Stephen W. Santoro, Andrew B. Martin, David S. King, Lei Wang, and Peter G. Schultz. (aug) 2002. “Addition Of P-Azido-L-Phenylalanine To The Genetic Code Of Escherichia Coli”. Journal Of The American Chemical Society 124: 9026-9027. doi:10.1021/ja027007w.
RS/tRNA Usage Publications
J, Islam, Conroy P, Fercher C, Kim M, Yaari Z, Jones M, Bell Tdm, et al. (aug) 2023. “Design Of Polarity-Dependent Immunosensors Based On The Structural Analysis Of Engineered Antibodies”. Acs Chemical Biology 18. doi:10.1021/acschembio.3c00303.
Ad, Guo, and Chen Xh. 2023. “Genetically Encoded Noncanonical Amino Acids In Proteins To Investigate Lysine Benzoylation”. Methods In Molecular Biology (Clifton, N.j.) 2676. doi:10.1007/978-1-0716-3251-2_9.
Y, Wang, Zhang J, Han B, Tan L, Cai W, Li Y, Su Y, et al. (feb) 2023. “Noncanonical Amino Acids As Doubly Bio-Orthogonal Handles For One-Pot Preparation Of Protein Multiconjugates”. Nature Communications 14. doi:10.1038/s41467-023-36658-y.
X, Ma, Wei B, and Wang E. (dec) 2022. “Efficient Incorporation Of P-Azido-L-Phenylalanine Into The Protein Using Organic Solvents”. Protein Expression And Purification 200. doi:10.1016/j.pep.2022.106158.
Cms, Michielsen, van Aalen Ea, and Merkx M. (jun) 2022. “Ratiometric Bioluminescent Zinc Sensor Proteins To Quantify Serum And Intracellular Free Zn2”. Acs Chemical Biology 17. doi:10.1021/acschembio.2c00227.
RS/tRNA Pair Development Year
2002
ncAA(s) Incorporated
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
Y32T
E107N
D158P
I159L
L162Q
D286R
E107N
D158P
I159L
L162Q
D286R
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
It can be procured from AddGene #31186 under the name pEVOL-AzF. Note the Addgene entry states that an additional D286R mutation is present that was not mentioned in the original publication.
RS/tRNA Additional Notes
Using 1 mM pAzF in glycerol minimal media w/ leucine, led to high-fidelity production of myoglobin(2) at 2mg/L. MS confirmed the fidelity. Also used to put pAzF into GST at 5 different sites and one site near the dimer interface allowed 10-30% crosslinking when photoactivated.