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MmPylRS-WT

RS/tRNA Foundational Publication Support

Blight, Sherry K, Ross C Larue, Anirban Mahapatra, David G Longstaff, Edward Chang, Gang Zhao, Patrick T Kang, Kari B Green-Church, Michael K Chan, and Joseph A Krzycki. (2004) 2004. “Direct Charging Of Trna(Cua) With Pyrrolysine In Vitro And In Vivo.”. Nature 431 (7006): 333-5.
Li, Xin, Tomasz Fekner, Jennifer J Ottesen, and Michael K Chan. (2009) 2009. “A Pyrrolysine Analogue For Site-Specific Protein Ubiquitination.”. Angewandte Chemie (International Ed. In English) 48 (48): 9184-7. doi:10.1002/anie.200904472.
Crnković, Ana, Tateki Suzuki, Dieter Söll, and Noah M Reynolds. (2016) 2016. “Pyrrolysyl-Trna Synthetase, An Aminoacyl-Trna Synthetase For Genetic Code Expansion.”. Croatica Chemica Acta. Arhiv Za Kemiju 89 (2): 163-174. doi:10.5562/cca2825.
Gattner, Michael J, Milan Vrabel, and Thomas Carell. (2013) 2013. “Synthesis Of Ε-N-Propionyl-, Ε-N-Butyryl-, And Ε-N-Crotonyl-Lysine Containing Histone H3 Using The Pyrrolysine System.”. Chemical Communications (Cambridge, England) 49 (4): 379-81. doi:10.1039/c2cc37836a.
Meineke, Birthe, Johannes Heimgärtner, Jürgen Eirich, Michael Landreh, and Simon J Elsässer. (2020) 2020. “Site-Specific Incorporation Of Two Ncaas For Two-Color Bioorthogonal Labeling And Crosslinking Of Proteins On Live Mammalian Cells.”. Cell Reports 31 (12): 107811. doi:10.1016/j.celrep.2020.107811.

RS/tRNA Protocols and Structural Information

Kavran, Jennifer M, Sarath Gundllapalli, Patrick O'Donoghue, Markus Englert, Dieter Söll, and Thomas A Steitz. (2007) 2007. “Structure Of Pyrrolysyl-Trna Synthetase, An Archaeal Enzyme For Genetic Code Innovation.”. Proceedings Of The National Academy Of Sciences Of The United States Of America 104 (27): 11268-73.

RS/tRNA Usage Publications

Yu, Zhipeng, and Qing Lin. (2014) 2014. “Design Of Spiro[2.3]Hex-1-Ene, A Genetically Encodable Double-Strained Alkene For Superfast Photoclick Chemistry.”. Journal Of The American Chemical Society 136 (11): 4153-6. doi:10.1021/ja5012542.
RS/tRNA Pair Development Year
2009

ncAA(s) Incorporated

Pyrrolysine (Pyl)

Pyrrolysine
ncAA Utility
serves as a key component in "genetic code expansion" techniques

l-Cys-e-Lys

N6-L -Cysteinyl-L-lysine
ncAA Utility
Condenses with 2-cyanobenzothiazole for protein modification.
RS Organism of Origin
Methanosarcina mazei
Parent RS
Pyl RS
RS Mutations
N/A
tRNA Organism of Origin
Methanosarcina mazei
Parent tRNA
Pyl tRNA
tRNA Anticodon
CUA
RS/tRNA Availability
N/A
Used in what cell line?
BL21(DE3)
RS/tRNA Additional Notes
In BL21(DE3) cells, mmPylRS-WT was transformed in terrific broth media alongside an mCherry reporter in the presence or absence of varying concentration of D-Cys-e-Lys and L-Cys-e-Lys. Fluorescence was only observed in the presence of either substrate, with D-Cys-e-Lys exhibiting greater expression.

Fifth foundational paper shows efficient incorporation of AzeoK and also N-propargyl-Lys (ProK)