Oligonucleotide-directed peptide synthesis in a ribosome- and ribozyme-free system.
Peptide bond formation by the ribosome requires 23S rRNA and its interaction with the 3'-CCA end of tRNA. To investigate the possible evolutionary development of the peptidyl transfer reaction, we tried to obtain peptide bond formation without the ribosome or rRNA simply by using a piece of tRNA--an aminoacyl-minihelix--mixed with sequence-specific oligonucleotides that contained puromycin. Peptide bond formation was detected by gel electrophoresis, TLC analysis, and mass spectrometry. Peptide synthesis depended on sequence complementarity between the 3'-CCA sequence of the minihelix and the puromycin-bearing oligonucleotide. However, proximity of the reacting species was not by itself sufficient for peptide bond formation. In addition, imidazole as a catalyst was required. Its role may be similar to the recently proposed mechanism, wherein A2451 of 23S rRNA works as a general base. Thus, peptide bond formation can be achieved with a simple, minimized system that captures the essence of an interaction seen in the ribosome.
Tamura
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K
The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Schimmel
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P
D020029:Base Pairing; D007093:Imidazoles; D009841:Oligonucleotides / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis*; D016337:RNA, Catalytic; D012343:RNA, Transfer / Q000737:chemistry*; D012270:Ribosomes
D007093:Imidazoles; D009841:Oligonucleotides; D010455:Peptides; D016337:RNA, Catalytic; C029899:imidazole; D012343:RNA, Transfer