Toward a Traceless Tag for the Thiol‐Mediated Uptake of Proteins
Abstract The emergence of thiol‐mediated uptake (TMU) as a powerful strategy to penetrate cells calls for the development of practical small‐molecule TMU tools for traceless delivery. Toward this goal, esters are explored here as bioreversible linkers between dynamic covalent cascade exchangers acco...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley-VCH
2023-11-01
|
| Series: | ChemistryEurope |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/ceur.202300029 |
| _version_ | 1827632035010707456 |
|---|---|
| author | Dr. John R. J. Maynard Dr. Saidbakhrom Saidjalolov Marie‐Claire Velluz Stefania Vossio Prof. Charlotte Aumeier Dr. Dimitri Moreau Dr. Naomi Sakai Prof. Stefan Matile |
| author_facet | Dr. John R. J. Maynard Dr. Saidbakhrom Saidjalolov Marie‐Claire Velluz Stefania Vossio Prof. Charlotte Aumeier Dr. Dimitri Moreau Dr. Naomi Sakai Prof. Stefan Matile |
| author_sort | Dr. John R. J. Maynard |
| collection | DOAJ |
| description | Abstract The emergence of thiol‐mediated uptake (TMU) as a powerful strategy to penetrate cells calls for the development of practical small‐molecule TMU tools for traceless delivery. Toward this goal, esters are explored here as bioreversible linkers between dynamic covalent cascade exchangers accounting for TMU and the protein of interest (POI). The method relies on α‐aryl‐α‐diazoamides that react with carboxylic acids of the POI to form esters that can be enzymatically hydrolyzed inside cells to release the native POI. A two‐step protocol is established for bioreversible conjugation of TMU tags to the POI. Despite the small number of tags attached to POIs to prevent isoelectric precipitation, POIs with traceless TMU tags are shown to efficiently enter cells not only in 2D culture but also in 3D spheroids mimicking deep tissue, confirming a key advantage of TMU. Uptake inhibition by various thiol‐reactive agents confirms the participation of cell‐surface thiols in cell penetration, i. e., the occurrence of TMU. |
| first_indexed | 2024-03-09T14:29:29Z |
| format | Article |
| id | doaj.art-219c132bebe948e7aa7e86e14fa9dfb3 |
| institution | Directory Open Access Journal |
| issn | 2751-4765 |
| language | English |
| last_indexed | 2024-03-09T14:29:29Z |
| publishDate | 2023-11-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | ChemistryEurope |
| spelling | doaj.art-219c132bebe948e7aa7e86e14fa9dfb32023-11-28T02:09:21ZengWiley-VCHChemistryEurope2751-47652023-11-0113n/an/a10.1002/ceur.202300029Toward a Traceless Tag for the Thiol‐Mediated Uptake of ProteinsDr. John R. J. Maynard0Dr. Saidbakhrom Saidjalolov1Marie‐Claire Velluz2Stefania Vossio3Prof. Charlotte Aumeier4Dr. Dimitri Moreau5Dr. Naomi Sakai6Prof. Stefan Matile7Department of Organic Chemistry University of Geneva 1211 Geneva SwitzerlandDepartment of Organic Chemistry University of Geneva 1211 Geneva SwitzerlandDepartment of Biochemistry University of Geneva 1211 Geneva SwitzerlandACCESS Geneva University of Geneva 1211 Geneva SwitzerlandDepartment of Biochemistry University of Geneva 1211 Geneva SwitzerlandACCESS Geneva University of Geneva 1211 Geneva SwitzerlandDepartment of Organic Chemistry University of Geneva 1211 Geneva SwitzerlandDepartment of Organic Chemistry University of Geneva 1211 Geneva SwitzerlandAbstract The emergence of thiol‐mediated uptake (TMU) as a powerful strategy to penetrate cells calls for the development of practical small‐molecule TMU tools for traceless delivery. Toward this goal, esters are explored here as bioreversible linkers between dynamic covalent cascade exchangers accounting for TMU and the protein of interest (POI). The method relies on α‐aryl‐α‐diazoamides that react with carboxylic acids of the POI to form esters that can be enzymatically hydrolyzed inside cells to release the native POI. A two‐step protocol is established for bioreversible conjugation of TMU tags to the POI. Despite the small number of tags attached to POIs to prevent isoelectric precipitation, POIs with traceless TMU tags are shown to efficiently enter cells not only in 2D culture but also in 3D spheroids mimicking deep tissue, confirming a key advantage of TMU. Uptake inhibition by various thiol‐reactive agents confirms the participation of cell‐surface thiols in cell penetration, i. e., the occurrence of TMU.https://doi.org/10.1002/ceur.202300029bioconjugationbioreversible esterificationisoelectric precipitationprotein deliveryspheroidsthiol-mediated uptake |
| spellingShingle | Dr. John R. J. Maynard Dr. Saidbakhrom Saidjalolov Marie‐Claire Velluz Stefania Vossio Prof. Charlotte Aumeier Dr. Dimitri Moreau Dr. Naomi Sakai Prof. Stefan Matile Toward a Traceless Tag for the Thiol‐Mediated Uptake of Proteins ChemistryEurope bioconjugation bioreversible esterification isoelectric precipitation protein delivery spheroids thiol-mediated uptake |
| title | Toward a Traceless Tag for the Thiol‐Mediated Uptake of Proteins |
| title_full | Toward a Traceless Tag for the Thiol‐Mediated Uptake of Proteins |
| title_fullStr | Toward a Traceless Tag for the Thiol‐Mediated Uptake of Proteins |
| title_full_unstemmed | Toward a Traceless Tag for the Thiol‐Mediated Uptake of Proteins |
| title_short | Toward a Traceless Tag for the Thiol‐Mediated Uptake of Proteins |
| title_sort | toward a traceless tag for the thiol mediated uptake of proteins |
| topic | bioconjugation bioreversible esterification isoelectric precipitation protein delivery spheroids thiol-mediated uptake |
| url | https://doi.org/10.1002/ceur.202300029 |
| work_keys_str_mv | AT drjohnrjmaynard towardatracelesstagforthethiolmediateduptakeofproteins AT drsaidbakhromsaidjalolov towardatracelesstagforthethiolmediateduptakeofproteins AT marieclairevelluz towardatracelesstagforthethiolmediateduptakeofproteins AT stefaniavossio towardatracelesstagforthethiolmediateduptakeofproteins AT profcharlotteaumeier towardatracelesstagforthethiolmediateduptakeofproteins AT drdimitrimoreau towardatracelesstagforthethiolmediateduptakeofproteins AT drnaomisakai towardatracelesstagforthethiolmediateduptakeofproteins AT profstefanmatile towardatracelesstagforthethiolmediateduptakeofproteins |