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Enhanced Live‐Cell Delivery of Synthetic Proteins Assisted by Cell‐Penetrating Peptides Fused to DABCYL
Live‐cell delivery of a fully synthetic protein having selectivity towards a particular target is a promising approach with potential applications for basic research and therapeutics. Cell‐penetrating peptides (CPPs) allow the cellular delivery of proteins but mostly result in endosomal entrapment,...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8048964/ https://www.ncbi.nlm.nih.gov/pubmed/33615660 http://dx.doi.org/10.1002/anie.202016208 |
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author | Mandal, Shaswati Mann, Guy Satish, Gandhesiri Brik, Ashraf |
author_facet | Mandal, Shaswati Mann, Guy Satish, Gandhesiri Brik, Ashraf |
author_sort | Mandal, Shaswati |
collection | PubMed |
description | Live‐cell delivery of a fully synthetic protein having selectivity towards a particular target is a promising approach with potential applications for basic research and therapeutics. Cell‐penetrating peptides (CPPs) allow the cellular delivery of proteins but mostly result in endosomal entrapment, leading to lack of bioavailability. Herein, we report the design and synthesis of a CPP fused to 4‐((4‐(dimethylamino)phenyl)azo)benzoic acid (DABCYL) to enhance cellular uptake of fluorescently labelled synthetic protein analogues in low micromolar concentration. The attachment of cyclic deca‐arginine (cR10) modified with a single lysine linked to DABCYL to synthetic ubiquitin (Ub) and small ubiquitin‐like modifier‐2 (SUMO‐2) scaffolds resulted in a threefold higher uptake efficacy in live cells compared to the unmodified cR10. We could also achieve cR10DABCYL‐assisted delivery of Ub and a Ub variant (Ubv) based activity‐based probes for functional studies of deubiquitinases in live cells. |
format | Online Article Text |
id | pubmed-8048964 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-80489642021-04-20 Enhanced Live‐Cell Delivery of Synthetic Proteins Assisted by Cell‐Penetrating Peptides Fused to DABCYL Mandal, Shaswati Mann, Guy Satish, Gandhesiri Brik, Ashraf Angew Chem Int Ed Engl Research Articles Live‐cell delivery of a fully synthetic protein having selectivity towards a particular target is a promising approach with potential applications for basic research and therapeutics. Cell‐penetrating peptides (CPPs) allow the cellular delivery of proteins but mostly result in endosomal entrapment, leading to lack of bioavailability. Herein, we report the design and synthesis of a CPP fused to 4‐((4‐(dimethylamino)phenyl)azo)benzoic acid (DABCYL) to enhance cellular uptake of fluorescently labelled synthetic protein analogues in low micromolar concentration. The attachment of cyclic deca‐arginine (cR10) modified with a single lysine linked to DABCYL to synthetic ubiquitin (Ub) and small ubiquitin‐like modifier‐2 (SUMO‐2) scaffolds resulted in a threefold higher uptake efficacy in live cells compared to the unmodified cR10. We could also achieve cR10DABCYL‐assisted delivery of Ub and a Ub variant (Ubv) based activity‐based probes for functional studies of deubiquitinases in live cells. John Wiley and Sons Inc. 2021-02-22 2021-03-22 /pmc/articles/PMC8048964/ /pubmed/33615660 http://dx.doi.org/10.1002/anie.202016208 Text en © 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) 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 | Research Articles Mandal, Shaswati Mann, Guy Satish, Gandhesiri Brik, Ashraf Enhanced Live‐Cell Delivery of Synthetic Proteins Assisted by Cell‐Penetrating Peptides Fused to DABCYL |
title | Enhanced Live‐Cell Delivery of Synthetic Proteins Assisted by Cell‐Penetrating Peptides Fused to DABCYL |
title_full | Enhanced Live‐Cell Delivery of Synthetic Proteins Assisted by Cell‐Penetrating Peptides Fused to DABCYL |
title_fullStr | Enhanced Live‐Cell Delivery of Synthetic Proteins Assisted by Cell‐Penetrating Peptides Fused to DABCYL |
title_full_unstemmed | Enhanced Live‐Cell Delivery of Synthetic Proteins Assisted by Cell‐Penetrating Peptides Fused to DABCYL |
title_short | Enhanced Live‐Cell Delivery of Synthetic Proteins Assisted by Cell‐Penetrating Peptides Fused to DABCYL |
title_sort | enhanced live‐cell delivery of synthetic proteins assisted by cell‐penetrating peptides fused to dabcyl |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8048964/ https://www.ncbi.nlm.nih.gov/pubmed/33615660 http://dx.doi.org/10.1002/anie.202016208 |
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