3-nitroTyr(5B) RS/tRNA(CUA) | GCE4All Knowledge Base

RS/tRNA Foundational Publication Support

RS/tRNA Usage Publications

Franco, Maria, Yaozu Ye, Christian A Refakis, Jessica L Feldman, Audrey L Stokes, Manuela Basso, Raquel de Mera, et al. (2013) 2013. “Nitration Of Hsp90 Induces Cell Death.”. Proceedings Of The National Academy Of Sciences Of The United States Of America 110 (12): E1102-11. doi:10.1073/pnas.1215177110.

Franco, Maria. (2015) 2015. “Nitration Of Hsp90 On Tyrosine 33 Regulates Mitochondrial Metabolism.”. The Journal Of Biological Chemistry 290 (31): 19055-66. doi:10.1074/jbc.M115.663278.

Gerding, Hanne, Christiaan Karreman, Andreas Daiber, Johannes Delp, Daniel Hammler, Martin Mex, Stefan Schildknecht, and Marcel Leist. (2019) 2019. “Reductive Modification Of Genetically Encoded 3-Nitrotyrosine Sites In Alpha Synuclein Expressed In E.coli.”. Redox Biology 26: 101251. doi:10.1016/j.redox.2019.101251.

DiDonato, Joseph A, Kulwant Aulak, Ying Huang, Matthew Wagner, Gary Gerstenecker, Celalettin Topbas, Valentin Gogonea, et al. (2014) 2014. “Site-Specific Nitration Of Apolipoprotein A-I At Tyrosine 166 Is Both Abundant Within Human Atherosclerotic Plaque And Dysfunctional.”. The Journal Of Biological Chemistry 289 (15): 10276-10292. doi:10.1074/jbc.M114.556506.

Zheng, Zhaopeng, Xuzhen Guo, Minling Yu, Xiaoyan Wang, Hongguang Lu, Fahui Li, and Jiangyun Wang. (2020) 2020. “Identification Of Human Ido1 Enzyme Activity By Using Genetically Encoded Nitrotyrosine.”. Chembiochem : A European Journal Of Chemical Biology 21 (11): 1593-1596. doi:10.1002/cbic.201900735.

Jandy, Megan, Asra Noor, Pascal Nelson, Cassandra Dennys, Isabella Karabinas, Jeanine Pestoni, Gautam Singh, et al. (2022) 2022. “Peroxynitrite Nitration Of Tyr 56 In Hsp90 Induces Pc12 Cell Death Through P2X7R-Dependent Pten Activation.”. Redox Biology 50: 102247. doi:10.1016/j.redox.2022.102247.

Logan, Isabelle, Kyle T Nguyen, Tilottama Chatterjee, Bhagyashree Manivannan, Ngozi Paul, Sharon Kim, Evelyn Sixta, et al. (2024) 2024. “Selective Nitration Of Hsp90 Acts As A Metabolic Switch Promoting Tumor Cell Proliferation.”. Redox Biology 75: 103249. doi:10.1016/j.redox.2024.103249.

Porter, Joseph J, Hyo Sang Jang, Mohammad Mahfuzul Haque, Dennis J Stuehr, and Ryan A Mehl. (2020) 2020. “Tyrosine Nitration On Calmodulin Enhances Calcium-Dependent Association And Activation Of Nitric-Oxide Synthase.”. The Journal Of Biological Chemistry 295 (8): 2203-2211. doi:10.1074/jbc.RA119.010999.

RS/tRNA Pair Development Year

2014

ncAA(s) Incorporated

RS Organism of Origin

Methanocaldococcus jannaschii

Parent RS

Tyr RS

RS Mutations

Y32H
H70C
D158S
I159A
L162R

tRNA Organism of Origin

Methanocaldococcus jannaschii

Parent tRNA

Tyr tRNA

tRNA Anticodon

CUA

Other tRNA Mutations

C17A
U17aG
U20C
G37A
U47G

Multiple tRNAs?

2016 paper showed that for this RS and also other MjTyr ncAA RSs, a tRNA having the CUA anticodon with only G37A as an additional mutation actually performed better than did the orthogonal tRNA containing the full set of 5 additional mutations listed above.

RS/tRNA Availability

RS and tRNA available as Addgene Plasmid #85498

Used in what cell line?

DH10B

Top10

RS/tRNA Additional Notes

About 10x more efficient than the first generation 3-nitroTyr RS, yielding high fidelity incorporation into sfGFP(150) using 1 mM ncAA with a yield of ~175 mg/L, representing about a 20% efficiency (relative to WT-sfGFP). Also yielded high fidelity incorporation into multiple sites in HSP90.

Crystal structures of the 5B 3-nitroTyr RS are available (PDB IDs: 4NDA, 4ND6 and 4ND7)