RS/tRNA Foundational Publication Support
Stokes, Audrey L, Shigeki J Miyake-Stoner, Jennifer C Peeler, Duy P Nguyen, Robert P Hammer, and Ryan A Mehl. (2009) 2009. “Enhancing The Utility Of Unnatural Amino Acid Synthetases By Manipulating Broad Substrate Specificity.”. Molecular Biosystems 5 (9): 1032-8. doi:10.1039/b904032c.
RS/tRNA Protocols and Structural Information
Peeler, Jennifer C, and Ryan A Mehl. (2012) 2012. “Site-Specific Incorporation Of Unnatural Amino Acids As Probes For Protein Conformational Changes.”. Methods In Molecular Biology (Clifton, N.j.) 794: 125-34. doi:10.1007/978-1-61779-331-8_8.
RS/tRNA Pair Development Year
2009
ncAA(s) Incorporated
4-Fluoro-p-benzoyl-L-phenylalanine(4F-Bpa)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Demonstration that one synthetase can have a rationally designed broad scope
Thyronine
ncAA Structure (png, jpg, jpeg)
ncAA Utility
n/a
p-benzoyl-L-phenylalanine (Bpa)
ncAA Structure (png, jpg, jpeg)
ncAA Utility
Used as a photocrosslinker, allowing for crosslinking and bioorthogonal ligation of protein.
RS Organism of Origin
Parent RS
RS Mutations
A31V
Y32G
E107P
D158S
I159S
Y32G
E107P
D158S
I159S
tRNA Organism of Origin
Parent tRNA
tRNA Anticodon
CUA
Other tRNA Mutations
C17A
U17aG
U20C
G37A
U47G
U17aG
U20C
G37A
U47G
RS/tRNA Availability
N/A
Used in what cell line?
RS/tRNA Additional Notes
In DH10B cells, with 1 mM 4F-Bpa or 1 mM Thyronine yielded ~1000 mg/L sfGFP(150). With 1 mM Bpa yield was ~850 mg/L, more than double the yield produced by the original BpaRS synthetase. for comparison, sfGFP wild-type yielded ~1800 mg/L.