Flexizyme as a versatile tRNA acylation catalyst and the application for translation.
Here we describe a de novo tRNA acylation system consisting of artificial ribozymes. This system, the flexizyme (Fx) system, allows for the preparation of acyl-tRNAs with almost no limitation on the use of a variety of amino/hydroxy acids and tRNAs. To demonstrate its utility, we have carried out protein synthesis involving site-specific incorporation of nonnatural amino and hydroxy acids via amber or programmed frame-shift suppressions. We have also demonstrated peptide synthesis involving multiple nonnatural amino acids via sense codon suppression. The combination of the Fx system and appropriate cell-free translation is a powerful and flexible tool for mRNA-encoded synthesis of nonnatural polypeptides.
Murakami
Hiroshi
H
Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Ohta
Atushi
A
Goto
Yuki
Y
Sako
Yusuke
Y
Suga
Hiroaki
H
D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism; D049148:Aminoacylation; D002384:Catalysis; D006880:Hydroxy Acids / Q000378:metabolism; D010452:Peptide Biosynthesis; D010455:Peptides / Q000737:chemistry; D014176:Protein Biosynthesis; D016337:RNA, Catalytic / Q000737:chemistry / Q000378:metabolism*; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
D000596:Amino Acids; D006880:Hydroxy Acids; D010455:Peptides; D016337:RNA, Catalytic; D012343:RNA, Transfer