RS/tRNA Foundational Publication Support
Furman, Jennifer L, Mingchao Kang, Seihyun Choi, Yu Cao, Erik D Wold, Sophie B Sun, Vaughn Smider V, Peter G Schultz, and Chan Hyuk Kim. (2014) 2014. “A Genetically Encoded Aza-Michael Acceptor For Covalent Cross-Linking Of Protein-Receptor Complexes.”. Journal Of The American Chemical Society 136 (23): 8411-7. doi:10.1021/ja502851h.
RS/tRNA Pair Development Year
2014
ncAA(s) Incorporated
p-Vinylsulfonamido (S)-L-phenylalanine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Reactive handle for Michael additions, cross-linker
RS Organism of Origin
Parent RS
RS Mutations
T32G
L65T
F108H
Q109G
D158G
I159L
L162Q
D286R
L65T
F108H
Q109G
D158G
I159L
L162Q
D286R
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
RS/tRNA Availability
N/A
Used in what cell line?
RS/tRNA Additional Notes
In BL21(DE3) cells, pET101-GFP-Tyr151TAG-His6 and pEVOL-VSFRS/tRNATyr were co-expressed in the presence and absence of 5mM p-Vinylsulfonamido (S)-L-phenylalanine (VSF), with incorporation confirmed by ESI-MS. The observed mass however did not line up with the expected mass of the mutant protein, which was explained as a GSH conjugate.
In TB media, Herceptin Fab-Tyr92TAG was expressed in the presence of VSFRS and 1mM supplemented VSF, resulting in a yield of 4mg/L mutant protein. ESI-MS characterization confirmed incorporation however a GSH conjugate was once again observed.
In TB media, Herceptin Fab-Tyr92TAG was expressed in the presence of VSFRS and 1mM supplemented VSF, resulting in a yield of 4mg/L mutant protein. ESI-MS characterization confirmed incorporation however a GSH conjugate was once again observed.