Charging of tRNA with non-natural amino acids at high pressure.
We show a simple and reliable method of tRNA aminoacylation with natural, as well as non-natural, amino acids at high pressure. Such specific and noncognate tRNAs can be used as valuable substrates for protein engineering. Aminoacylation yield at high pressure depends on the chemical nature of the amino acid used and it is up to 10%. Using CoA, which carries two potentially reactive groups -SH and -OH, as a model compound we showed that at high pressure amino acid is bound preferentially to the hydroxyl group of the terminal ribose ring.
Giel-Pietraszuk
Malgorzata
M
Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznan, Poland.
Barciszewski
Jan
J
D000596:Amino Acids / Q000737:chemistry*; D001671:Biochemistry / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003062:Codon; D004926:Escherichia coli / Q000378:metabolism; D005658:Fungi / Q000378:metabolism; D013058:Mass Spectrometry; D008956:Models, Chemical; D011312:Pressure; D015202:Protein Engineering / Q000379:methods*; D012343:RNA, Transfer / Q000737:chemistry*; D013997:Time Factors; D046249:Transfer RNA Aminoacylation
D000596:Amino Acids; D003062:Codon; D012343:RNA, Transfer