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Semi-Synthesis and Analysis of Chemically Modified Zif268 Zinc-Finger Domains

Total synthesis of proteins can be challenging despite assembling techniques, such as native chemical ligation (NCL) and expressed protein ligation (EPL). Especially, the combination of recombinant protein expression and chemically addressable solid-phase peptide synthesis (SPPS) is well suited for...

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Autores principales: Fehr, Friederike, Nadler, André, Brodhun, Florian, Feussner, Ivo, Diederichsen, Ulf
Formato: Online Artículo Texto
Lenguaje:English
Publicado: WILEY-VCH Verlag 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3922437/
https://www.ncbi.nlm.nih.gov/pubmed/24551489
http://dx.doi.org/10.1002/open.201100002
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author Fehr, Friederike
Nadler, André
Brodhun, Florian
Feussner, Ivo
Diederichsen, Ulf
author_facet Fehr, Friederike
Nadler, André
Brodhun, Florian
Feussner, Ivo
Diederichsen, Ulf
author_sort Fehr, Friederike
collection PubMed
description Total synthesis of proteins can be challenging despite assembling techniques, such as native chemical ligation (NCL) and expressed protein ligation (EPL). Especially, the combination of recombinant protein expression and chemically addressable solid-phase peptide synthesis (SPPS) is well suited for the redesign of native protein structures. Incorporation of analytical probes and artificial amino acids into full-length natural protein domains, such as the sequence-specific DNA binding zinc-finger motifs, are of interest combining selective DNA recognition and artificial function. The semi-synthesis of the natural 90 amino acid long sequence of the zinc-finger domain of Zif268 is described including various chemically modified constructs. Our approach offers the possibility to exchange any amino acid within the third zinc finger. The realized modifications of the natural sequence include point mutations, attachment of a fluorophore, and the exchange of amino acids at different positions in the zinc finger by artificial amino acids to create additional metal binding sites. The individual constructs were analyzed by circular dichroism (CD) spectroscopy with respect to the integrity of the zinc-finger fold and DNA binding.
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spelling pubmed-39224372014-02-18 Semi-Synthesis and Analysis of Chemically Modified Zif268 Zinc-Finger Domains Fehr, Friederike Nadler, André Brodhun, Florian Feussner, Ivo Diederichsen, Ulf ChemistryOpen Full Papers Total synthesis of proteins can be challenging despite assembling techniques, such as native chemical ligation (NCL) and expressed protein ligation (EPL). Especially, the combination of recombinant protein expression and chemically addressable solid-phase peptide synthesis (SPPS) is well suited for the redesign of native protein structures. Incorporation of analytical probes and artificial amino acids into full-length natural protein domains, such as the sequence-specific DNA binding zinc-finger motifs, are of interest combining selective DNA recognition and artificial function. The semi-synthesis of the natural 90 amino acid long sequence of the zinc-finger domain of Zif268 is described including various chemically modified constructs. Our approach offers the possibility to exchange any amino acid within the third zinc finger. The realized modifications of the natural sequence include point mutations, attachment of a fluorophore, and the exchange of amino acids at different positions in the zinc finger by artificial amino acids to create additional metal binding sites. The individual constructs were analyzed by circular dichroism (CD) spectroscopy with respect to the integrity of the zinc-finger fold and DNA binding. WILEY-VCH Verlag 2012-02 2012-01-02 /pmc/articles/PMC3922437/ /pubmed/24551489 http://dx.doi.org/10.1002/open.201100002 Text en Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim http://creativecommons.org/licenses/by/3.0/ This is an open access article under the terms of the Creative Commons Attribution Non-Commercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Full Papers
Fehr, Friederike
Nadler, André
Brodhun, Florian
Feussner, Ivo
Diederichsen, Ulf
Semi-Synthesis and Analysis of Chemically Modified Zif268 Zinc-Finger Domains
title Semi-Synthesis and Analysis of Chemically Modified Zif268 Zinc-Finger Domains
title_full Semi-Synthesis and Analysis of Chemically Modified Zif268 Zinc-Finger Domains
title_fullStr Semi-Synthesis and Analysis of Chemically Modified Zif268 Zinc-Finger Domains
title_full_unstemmed Semi-Synthesis and Analysis of Chemically Modified Zif268 Zinc-Finger Domains
title_short Semi-Synthesis and Analysis of Chemically Modified Zif268 Zinc-Finger Domains
title_sort semi-synthesis and analysis of chemically modified zif268 zinc-finger domains
topic Full Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3922437/
https://www.ncbi.nlm.nih.gov/pubmed/24551489
http://dx.doi.org/10.1002/open.201100002
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