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Four-base codon mediated mRNA display to construct peptide libraries that contain multiple nonnatural amino acids
In vitro selection and directed evolution of peptides from mRNA display are powerful strategies to find novel peptide ligands that bind to target biomolecules. In this study, we expanded the mRNA display method to include multiple nonnatural amino acids by introducing three different four-base codon...
Autores principales: | , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2006
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1325208/ https://www.ncbi.nlm.nih.gov/pubmed/16397292 http://dx.doi.org/10.1093/nar/gnj003 |
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author | Muranaka, Norihito Hohsaka, Takahiro Sisido, Masahiko |
author_facet | Muranaka, Norihito Hohsaka, Takahiro Sisido, Masahiko |
author_sort | Muranaka, Norihito |
collection | PubMed |
description | In vitro selection and directed evolution of peptides from mRNA display are powerful strategies to find novel peptide ligands that bind to target biomolecules. In this study, we expanded the mRNA display method to include multiple nonnatural amino acids by introducing three different four-base codons at a randomly selected single position on the mRNA. Another nonnatural amino acid may be introduced by suppressing an amber codon that may appear from a (NNK)(n) nucleotide sequence on the mRNA. The mRNA display was expressed in an Escherichia coli in vitro translation system in the presence of three types of tRNAs carrying different four-base anticodons and a tRNA carrying an amber anticodon, the tRNAs being chemically aminoacylated with different nonnatural amino acids. The complexity of the starting mRNA-displayed peptide library was estimated to be 1.1 × 10(12) molecules. The effectiveness of the four-base codon mediated mRNA display method was demonstrated in the selection of biocytin-containing peptides on streptavidin-coated beads. Moreover, a novel streptavidin-binding nonnatural peptide containing benzoylphenylalanine was obtained from the nonnatural peptide library. The nonnatural peptide library from the four-base codon mediated mRNA display provides much wider functional and structural diversity than conventional peptide libraries that are constituted from 20 naturally occurring amino acids. |
format | Text |
id | pubmed-1325208 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2006 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-13252082006-01-10 Four-base codon mediated mRNA display to construct peptide libraries that contain multiple nonnatural amino acids Muranaka, Norihito Hohsaka, Takahiro Sisido, Masahiko Nucleic Acids Res Methods Online In vitro selection and directed evolution of peptides from mRNA display are powerful strategies to find novel peptide ligands that bind to target biomolecules. In this study, we expanded the mRNA display method to include multiple nonnatural amino acids by introducing three different four-base codons at a randomly selected single position on the mRNA. Another nonnatural amino acid may be introduced by suppressing an amber codon that may appear from a (NNK)(n) nucleotide sequence on the mRNA. The mRNA display was expressed in an Escherichia coli in vitro translation system in the presence of three types of tRNAs carrying different four-base anticodons and a tRNA carrying an amber anticodon, the tRNAs being chemically aminoacylated with different nonnatural amino acids. The complexity of the starting mRNA-displayed peptide library was estimated to be 1.1 × 10(12) molecules. The effectiveness of the four-base codon mediated mRNA display method was demonstrated in the selection of biocytin-containing peptides on streptavidin-coated beads. Moreover, a novel streptavidin-binding nonnatural peptide containing benzoylphenylalanine was obtained from the nonnatural peptide library. The nonnatural peptide library from the four-base codon mediated mRNA display provides much wider functional and structural diversity than conventional peptide libraries that are constituted from 20 naturally occurring amino acids. Oxford University Press 2006 2006-01-05 /pmc/articles/PMC1325208/ /pubmed/16397292 http://dx.doi.org/10.1093/nar/gnj003 Text en © The Author 2006. Published by Oxford University Press. All rights reserved |
spellingShingle | Methods Online Muranaka, Norihito Hohsaka, Takahiro Sisido, Masahiko Four-base codon mediated mRNA display to construct peptide libraries that contain multiple nonnatural amino acids |
title | Four-base codon mediated mRNA display to construct peptide libraries that contain multiple nonnatural amino acids |
title_full | Four-base codon mediated mRNA display to construct peptide libraries that contain multiple nonnatural amino acids |
title_fullStr | Four-base codon mediated mRNA display to construct peptide libraries that contain multiple nonnatural amino acids |
title_full_unstemmed | Four-base codon mediated mRNA display to construct peptide libraries that contain multiple nonnatural amino acids |
title_short | Four-base codon mediated mRNA display to construct peptide libraries that contain multiple nonnatural amino acids |
title_sort | four-base codon mediated mrna display to construct peptide libraries that contain multiple nonnatural amino acids |
topic | Methods Online |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1325208/ https://www.ncbi.nlm.nih.gov/pubmed/16397292 http://dx.doi.org/10.1093/nar/gnj003 |
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