Multiple incorporation of non-natural amino acids into a single protein using tRNAs with non-standard structures.
The ability to introduce non-natural amino acids into proteins opens up new vistas for the study of protein structure and function. This approach requires suppressor tRNAs that deliver the non-natural amino acid to a ribosome associated with an mRNA containing an expanded codon. The suppressor tRNAs must be absolutely protected from aminoacylation by any of the aminoacyl-tRNA synthetases in the protein synthesizing system, or a natural amino acid will be incorporated instead of the non-natural amino acid. Here, we found that some tRNAs with non-standard structures could work as efficient four-base suppressors fulfilling the above orthogonal conditions. Using these tRNAs, we successfully demonstrated incorporation of three different non-natural amino acids into a single protein.
Ohtsuki
Takashi
T
Department of Bioscience and Biotechnology, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
Manabe
Taishi
T
Sisido
Masahiko
M
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry*; D049148:Aminoacylation; D000926:Anticodon / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D003062:Codon / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000138:chemical synthesis* / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism*; D012346:RNA, Transfer, Amino Acyl / Q000138:chemical synthesis; D013489:Suppression, Genetic
D000596:Amino Acids; D000926:Anticodon; D003062:Codon; D011506:Proteins; D012333:RNA, Messenger; D012346:RNA, Transfer, Amino Acyl; D012343:RNA, Transfer