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Kinetic analysis of the leucyl/phenylalanyl-tRNA-protein transferase with acceptor peptides possessing different N-terminal penultimate residues()
The introduction of non-natural amino acids at the N-terminus of peptides/proteins using leucyl/phenylalanyl-tRNA-protein transferase (L/F-transferase) is a useful technique for protein engineering. To accelerate the chemoenzymatic reaction, here we systematically optimized the N-terminal penultimat...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3722611/ https://www.ncbi.nlm.nih.gov/pubmed/23905007 http://dx.doi.org/10.1016/j.fob.2013.06.001 |
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author | Kawaguchi, Jun Maejima, Kumino Kuroiwa, Hiroyuki Taki, Masumi |
author_facet | Kawaguchi, Jun Maejima, Kumino Kuroiwa, Hiroyuki Taki, Masumi |
author_sort | Kawaguchi, Jun |
collection | PubMed |
description | The introduction of non-natural amino acids at the N-terminus of peptides/proteins using leucyl/phenylalanyl-tRNA-protein transferase (L/F-transferase) is a useful technique for protein engineering. To accelerate the chemoenzymatic reaction, here we systematically optimized the N-terminal penultimate residue of the acceptor peptide. Positively charged, small, or hydrophilic amino acids at this position show positive effects for the reaction. Kinetic analysis of peptides possessing different penultimate residues suggests that the side chain of the residue affects peptide-binding affinity towards the L/F-transferase. These findings also provide biological insight into the effect of the penultimate amino acid on substrate specificity of natural proteins to be degraded via the N-end rule pathway. |
format | Online Article Text |
id | pubmed-3722611 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-37226112013-07-31 Kinetic analysis of the leucyl/phenylalanyl-tRNA-protein transferase with acceptor peptides possessing different N-terminal penultimate residues() Kawaguchi, Jun Maejima, Kumino Kuroiwa, Hiroyuki Taki, Masumi FEBS Open Bio Article The introduction of non-natural amino acids at the N-terminus of peptides/proteins using leucyl/phenylalanyl-tRNA-protein transferase (L/F-transferase) is a useful technique for protein engineering. To accelerate the chemoenzymatic reaction, here we systematically optimized the N-terminal penultimate residue of the acceptor peptide. Positively charged, small, or hydrophilic amino acids at this position show positive effects for the reaction. Kinetic analysis of peptides possessing different penultimate residues suggests that the side chain of the residue affects peptide-binding affinity towards the L/F-transferase. These findings also provide biological insight into the effect of the penultimate amino acid on substrate specificity of natural proteins to be degraded via the N-end rule pathway. Elsevier 2013-06-11 /pmc/articles/PMC3722611/ /pubmed/23905007 http://dx.doi.org/10.1016/j.fob.2013.06.001 Text en © 2013 The Authors http://creativecommons.org/licenses/BY-NC-ND/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License, which permits non-commercial use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Article Kawaguchi, Jun Maejima, Kumino Kuroiwa, Hiroyuki Taki, Masumi Kinetic analysis of the leucyl/phenylalanyl-tRNA-protein transferase with acceptor peptides possessing different N-terminal penultimate residues() |
title | Kinetic analysis of the leucyl/phenylalanyl-tRNA-protein transferase with acceptor peptides possessing different N-terminal penultimate residues() |
title_full | Kinetic analysis of the leucyl/phenylalanyl-tRNA-protein transferase with acceptor peptides possessing different N-terminal penultimate residues() |
title_fullStr | Kinetic analysis of the leucyl/phenylalanyl-tRNA-protein transferase with acceptor peptides possessing different N-terminal penultimate residues() |
title_full_unstemmed | Kinetic analysis of the leucyl/phenylalanyl-tRNA-protein transferase with acceptor peptides possessing different N-terminal penultimate residues() |
title_short | Kinetic analysis of the leucyl/phenylalanyl-tRNA-protein transferase with acceptor peptides possessing different N-terminal penultimate residues() |
title_sort | kinetic analysis of the leucyl/phenylalanyl-trna-protein transferase with acceptor peptides possessing different n-terminal penultimate residues() |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3722611/ https://www.ncbi.nlm.nih.gov/pubmed/23905007 http://dx.doi.org/10.1016/j.fob.2013.06.001 |
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