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Structure–Uptake Relationship Study of DABCYL Derivatives Linked to Cyclic Cell‐Penetrating Peptides for Live‐Cell Delivery of Synthetic Proteins

Modifying cyclic cell‐penetrating deca‐arginine (cR10) peptides with 4‐(4‐dimethylaminophenylazo)benzoic acid (DABCYL) improves the uptake efficiency of synthetic ubiquitin (Ub) cargoes into living cells. To probe the role of the DABCYL moiety, we performed time‐lapse microscopy and fluorescence lif...

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Autores principales: Saha, Abhishek, Mandal, Shaswati, Arafiles, Jan Vincent V., Gómez‐González, Jacobo, Hackenberger, Christian P. R., Brik, Ashraf
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9828537/
https://www.ncbi.nlm.nih.gov/pubmed/36004945
http://dx.doi.org/10.1002/anie.202207551
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author Saha, Abhishek
Mandal, Shaswati
Arafiles, Jan Vincent V.
Gómez‐González, Jacobo
Hackenberger, Christian P. R.
Brik, Ashraf
author_facet Saha, Abhishek
Mandal, Shaswati
Arafiles, Jan Vincent V.
Gómez‐González, Jacobo
Hackenberger, Christian P. R.
Brik, Ashraf
author_sort Saha, Abhishek
collection PubMed
description Modifying cyclic cell‐penetrating deca‐arginine (cR10) peptides with 4‐(4‐dimethylaminophenylazo)benzoic acid (DABCYL) improves the uptake efficiency of synthetic ubiquitin (Ub) cargoes into living cells. To probe the role of the DABCYL moiety, we performed time‐lapse microscopy and fluorescence lifetime imaging microscopy (FLIM) of fluorescent DABCYL‐R10 to evaluate the impact on cell entry by the formation of nucleation zones. Furthermore, we performed a structure–uptake relationship study with 13 DABCYL derivatives coupled to CPP to examine their effect on the cell‐uptake efficiency when conjugated to mono‐Ub through disulfide linkages. Our results show that through structure variations of the DABCYL moiety alone we could reach, at nanomolar concentration, an additional threefold increase in the cytosolic delivery of Ub, which will enable studies on various intracellular processes related to Ub signaling.
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spelling pubmed-98285372023-01-10 Structure–Uptake Relationship Study of DABCYL Derivatives Linked to Cyclic Cell‐Penetrating Peptides for Live‐Cell Delivery of Synthetic Proteins Saha, Abhishek Mandal, Shaswati Arafiles, Jan Vincent V. Gómez‐González, Jacobo Hackenberger, Christian P. R. Brik, Ashraf Angew Chem Int Ed Engl Research Articles Modifying cyclic cell‐penetrating deca‐arginine (cR10) peptides with 4‐(4‐dimethylaminophenylazo)benzoic acid (DABCYL) improves the uptake efficiency of synthetic ubiquitin (Ub) cargoes into living cells. To probe the role of the DABCYL moiety, we performed time‐lapse microscopy and fluorescence lifetime imaging microscopy (FLIM) of fluorescent DABCYL‐R10 to evaluate the impact on cell entry by the formation of nucleation zones. Furthermore, we performed a structure–uptake relationship study with 13 DABCYL derivatives coupled to CPP to examine their effect on the cell‐uptake efficiency when conjugated to mono‐Ub through disulfide linkages. Our results show that through structure variations of the DABCYL moiety alone we could reach, at nanomolar concentration, an additional threefold increase in the cytosolic delivery of Ub, which will enable studies on various intracellular processes related to Ub signaling. John Wiley and Sons Inc. 2022-10-19 2022-11-21 /pmc/articles/PMC9828537/ /pubmed/36004945 http://dx.doi.org/10.1002/anie.202207551 Text en © 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Research Articles
Saha, Abhishek
Mandal, Shaswati
Arafiles, Jan Vincent V.
Gómez‐González, Jacobo
Hackenberger, Christian P. R.
Brik, Ashraf
Structure–Uptake Relationship Study of DABCYL Derivatives Linked to Cyclic Cell‐Penetrating Peptides for Live‐Cell Delivery of Synthetic Proteins
title Structure–Uptake Relationship Study of DABCYL Derivatives Linked to Cyclic Cell‐Penetrating Peptides for Live‐Cell Delivery of Synthetic Proteins
title_full Structure–Uptake Relationship Study of DABCYL Derivatives Linked to Cyclic Cell‐Penetrating Peptides for Live‐Cell Delivery of Synthetic Proteins
title_fullStr Structure–Uptake Relationship Study of DABCYL Derivatives Linked to Cyclic Cell‐Penetrating Peptides for Live‐Cell Delivery of Synthetic Proteins
title_full_unstemmed Structure–Uptake Relationship Study of DABCYL Derivatives Linked to Cyclic Cell‐Penetrating Peptides for Live‐Cell Delivery of Synthetic Proteins
title_short Structure–Uptake Relationship Study of DABCYL Derivatives Linked to Cyclic Cell‐Penetrating Peptides for Live‐Cell Delivery of Synthetic Proteins
title_sort structure–uptake relationship study of dabcyl derivatives linked to cyclic cell‐penetrating peptides for live‐cell delivery of synthetic proteins
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9828537/
https://www.ncbi.nlm.nih.gov/pubmed/36004945
http://dx.doi.org/10.1002/anie.202207551
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