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Comparison of Functional Protein Transduction Domains Using the NEMO Binding Domain Peptide

Protein transduction domains (PTDs), both naturally occurring and synthetic, have been extensively utilized for intracellular delivery of biologically active molecules both in vitro and in vivo. However, most comparisons of transduction efficiency have been performed using fluorescent markers. To co...

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Detalles Bibliográficos
Autores principales: Khaja, Khaleel, Robbins, Paul
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3991022/
https://www.ncbi.nlm.nih.gov/pubmed/27713244
http://dx.doi.org/10.3390/ph3010110
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author Khaja, Khaleel
Robbins, Paul
author_facet Khaja, Khaleel
Robbins, Paul
author_sort Khaja, Khaleel
collection PubMed
description Protein transduction domains (PTDs), both naturally occurring and synthetic, have been extensively utilized for intracellular delivery of biologically active molecules both in vitro and in vivo. However, most comparisons of transduction efficiency have been performed using fluorescent markers. To compare efficiency of functional protein transduction, a peptide derived from IκB kinase ß (IKKß) that prevents formation of an active IKK complex was used as a biologically active cargo. This peptide, termed NEMO Binding Domain (NBD), is able to block activation of the transcriptional factor NF-κB by IKK, but not basal NF-κB activity. Our results demonstrate that Antp and Tat PTDs were most effective for delivery of NBD for inhibition of NF-κB activation compared to other PTD-NBD in both Hela and 293 cells, however, at higher concentrations (100 µM), the Antp-NBD as well as the FGF-NBD peptide caused significant cellular toxicity. In contrast to the cell culture results, delivery of NBD using 8K (octalysine) and 6R (six arginine) were the most effect in blocking inflammation following local, footpad delivery in a KLH-induced DTH murine model of inflammatory arthritis. These results demonstrate differences between PTDs for delivery of a functional cargo between cell types.
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spelling pubmed-39910222014-04-18 Comparison of Functional Protein Transduction Domains Using the NEMO Binding Domain Peptide Khaja, Khaleel Robbins, Paul Pharmaceuticals (Basel) Article Protein transduction domains (PTDs), both naturally occurring and synthetic, have been extensively utilized for intracellular delivery of biologically active molecules both in vitro and in vivo. However, most comparisons of transduction efficiency have been performed using fluorescent markers. To compare efficiency of functional protein transduction, a peptide derived from IκB kinase ß (IKKß) that prevents formation of an active IKK complex was used as a biologically active cargo. This peptide, termed NEMO Binding Domain (NBD), is able to block activation of the transcriptional factor NF-κB by IKK, but not basal NF-κB activity. Our results demonstrate that Antp and Tat PTDs were most effective for delivery of NBD for inhibition of NF-κB activation compared to other PTD-NBD in both Hela and 293 cells, however, at higher concentrations (100 µM), the Antp-NBD as well as the FGF-NBD peptide caused significant cellular toxicity. In contrast to the cell culture results, delivery of NBD using 8K (octalysine) and 6R (six arginine) were the most effect in blocking inflammation following local, footpad delivery in a KLH-induced DTH murine model of inflammatory arthritis. These results demonstrate differences between PTDs for delivery of a functional cargo between cell types. Molecular Diversity Preservation International 2010-01-08 /pmc/articles/PMC3991022/ /pubmed/27713244 http://dx.doi.org/10.3390/ph3010110 Text en © 2010 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland http://creativecommons.org/licenses/by/3.0/ This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Khaja, Khaleel
Robbins, Paul
Comparison of Functional Protein Transduction Domains Using the NEMO Binding Domain Peptide
title Comparison of Functional Protein Transduction Domains Using the NEMO Binding Domain Peptide
title_full Comparison of Functional Protein Transduction Domains Using the NEMO Binding Domain Peptide
title_fullStr Comparison of Functional Protein Transduction Domains Using the NEMO Binding Domain Peptide
title_full_unstemmed Comparison of Functional Protein Transduction Domains Using the NEMO Binding Domain Peptide
title_short Comparison of Functional Protein Transduction Domains Using the NEMO Binding Domain Peptide
title_sort comparison of functional protein transduction domains using the nemo binding domain peptide
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3991022/
https://www.ncbi.nlm.nih.gov/pubmed/27713244
http://dx.doi.org/10.3390/ph3010110
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