RS/tRNA Foundational Publication Support
Link, James, Mandy Vink, Nicholas Agard, Jennifer A Prescher, Carolyn R Bertozzi, and David A Tirrell. (2006) 2006. “Discovery Of Aminoacyl-Trna Synthetase Activity Through Cell-Surface Display Of Noncanonical Amino Acids.”. Proceedings Of The National Academy Of Sciences Of The United States Of America 103 (27): 10180-10185. doi:10.1073/pnas.0601167103.
Tanrikulu, Caglar, Emmanuelle Schmitt, Yves Mechulam, William A Goddard, and David A Tirrell. (2009) 2009. “Discovery Of Escherichia Coli Methionyl-Trna Synthetase Mutants For Efficient Labeling Of Proteins With Azidonorleucine In Vivo.”. Proceedings Of The National Academy Of Sciences Of The United States Of America 106 (36): 15285-90. doi:10.1073/pnas.0905735106.
RS/tRNA Pair Development Year
2006
ncAA(s) Incorporated
Azidonorleucine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Biorthogonal Ligation, Click Chemistry labeling with fluorescence or affinity labels. Specifically used for cell-specific proteomics studies using "bioorthogonal noncanonical amino acid tagging" (BONCAT) proteomics. Contrasts with Azidohomoalanine which does not allow cell-specific studies.
RS Organism of Origin
Parent RS
RS Mutations
L13G
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
RS/tRNA Availability
N/A for the RS; the tRNA is endogenous to E. coli
Used in what cell line?
RS/tRNA Additional Notes
Met auxotroph (M15MA) E coli containing MetRS-L13G exhibited near complete replacement of Met by Anl when protein synthesis occurred in a Met-free minimal medium supplemented by 1.0 mM Anl.
In 2009, MetRS-L13G was found to activate Met better than Anl by a factor of 30, and then the NLL MetRS variant (L13N, Y260L, H301L) was selected and shown to be much better, capable of inserting Anl even in the presence of normal levels of Met.
In 2009, MetRS-L13G was found to activate Met better than Anl by a factor of 30, and then the NLL MetRS variant (L13N, Y260L, H301L) was selected and shown to be much better, capable of inserting Anl even in the presence of normal levels of Met.