RS/tRNA Foundational Publication Support
Brustad, Eric, Mark L Bushey, Ansgar Brock, Johnathan Chittuluru, and Peter G Schultz. (2008) 2008. “A Promiscuous Aminoacyl-Trna Synthetase That Incorporates Cysteine, Methionine, And Alanine Homologs Into Proteins.”. Bioorganic & Medicinal Chemistry Letters 18 (22): 6004-6. doi:10.1016/j.bmcl.2008.09.050.
RS/tRNA Pair Development Year
2008
ncAA(s) Incorporated
6-Sulfanyl-L-norleucine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
n/a
L-dihomomethionine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
n/a
2-aminooctanoic acid
ncAA Structure (png, jpg, jpeg)
ncAA Utility
n/a
RS Organism of Origin
Parent RS
RS Mutations
M40I
T499I
T527A
H537G
T252R
T247V
T248V
T499I
T527A
H537G
T252R
T247V
T248V
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
RS/tRNA Availability
N/A
Used in what cell line?
RS/tRNA Additional Notes
In SCY4 Yeast cells, EcLRS-DHE6 was co-transformed with pC1hSOD33TAG in the presence of 1mM L-dihomomethionine or 1mM 2-aminooctanoic acid, yielding 0.5mg/L and 0.3mg/L respectively. In the absence of either ncAA only low background presence of hSOD was observed, and MALDI-TOF mass spectrometry was used to confirm the incorporation.
EcLRS-DHE6 was spotted onto ura-agar plates supplemented with L-dihomomethionine, 2-aminooctanoic acid, 6-Sulfanyl-L-norleucine, as well as other ncAA, however these 3 were the only ones with the RS was effective for. Colonies were observed after 48 hours with the distinguishing factor being that these 3 amino acids had six carbon side chains. L-dihomomethionine and 2-aminooctanoic acid were introduced into hSOD-
Trp33TAG with incorporation again confirmed by MALDI-TOF spectrometry. 6-Sulfanyl-L-norleucine incorporation into hSOD was confirmed via tryptic-digest and subsequent LC-MS/MS.
EcLRS-DHE6 was spotted onto ura-agar plates supplemented with L-dihomomethionine, 2-aminooctanoic acid, 6-Sulfanyl-L-norleucine, as well as other ncAA, however these 3 were the only ones with the RS was effective for. Colonies were observed after 48 hours with the distinguishing factor being that these 3 amino acids had six carbon side chains. L-dihomomethionine and 2-aminooctanoic acid were introduced into hSOD-
Trp33TAG with incorporation again confirmed by MALDI-TOF spectrometry. 6-Sulfanyl-L-norleucine incorporation into hSOD was confirmed via tryptic-digest and subsequent LC-MS/MS.