Researchers have developed a technology to precisely incorporate a range of non-canonical amino acids into proteins made in eukaryotes. The research was done by chemists at Boston College and reported in Cell Chemical Biology.
The potential benefits of an aminoacyl-tRNA synthetase/tRNA pair for incorporating non-canonical amino acids into proteins had previously been identified, but technical difficulties in developing such a method has limited its widespread application. In the current study, the researchers overcame those limitations by developing a novel strain of E. coli to facilitate engineering of the bacteria-derived aminoacyl-tRNA synthetase/tRNA pair.
"Creating this novel E. coli strain required substituting its native aminoacyl-tRNA synthetase/tRNA pair with a counterpart from a different organism, which we anticipated would be very difficult," said Assistant Professor of Chemistry Abhishek Chatterjee. "But it turned out to be quite feasible. That opens up this complete technology."
The researchers created this technique to enable better understanding of the process that guides protein production and function by allowing introduction of probes to monitor protein-coding genes. Previous attempts to introduce such probes had been difficult, as the process often damages the target protein.
The aminoacyl-tRNA synthetase/tRNA pair in the engineered gene provides a site for introduction of non-canonical amino acids, which can be loaded with such probes or used to manipulate a protein’s function. The researchers believe this tool will open the door to more in-depth understanding of proteins and their specific roles in Eukaryotes.
Image: Boston College chemists have developed a new, bacteria-derived technology to incorporate non-canonical amino acids into proteins of a broad class of organisms, including humans. Image courtesy of Cell Chemical Biology.