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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...

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Autores principales: Muranaka, Norihito, Hohsaka, Takahiro, Sisido, Masahiko
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2006
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.
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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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