Cargando…

Rapid labelling and covalent inhibition of intracellular native proteins using ligand-directed N-acyl-N-alkyl sulfonamide

Selective modification of native proteins in live cells is one of the central challenges in recent chemical biology. As a unique bioorthogonal approach, ligand-directed chemistry recently emerged, but the slow kinetics limits its scope. Here we successfully overcome this obstacle using N-acyl-N-alky...

Descripción completa

Detalles Bibliográficos
Autores principales: Tamura, Tomonori, Ueda, Tsuyoshi, Goto, Taiki, Tsukidate, Taku, Shapira, Yonatan, Nishikawa, Yuki, Fujisawa, Alma, Hamachi, Itaru
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5951806/
https://www.ncbi.nlm.nih.gov/pubmed/29760386
http://dx.doi.org/10.1038/s41467-018-04343-0
_version_ 1783323072449740800
author Tamura, Tomonori
Ueda, Tsuyoshi
Goto, Taiki
Tsukidate, Taku
Shapira, Yonatan
Nishikawa, Yuki
Fujisawa, Alma
Hamachi, Itaru
author_facet Tamura, Tomonori
Ueda, Tsuyoshi
Goto, Taiki
Tsukidate, Taku
Shapira, Yonatan
Nishikawa, Yuki
Fujisawa, Alma
Hamachi, Itaru
author_sort Tamura, Tomonori
collection PubMed
description Selective modification of native proteins in live cells is one of the central challenges in recent chemical biology. As a unique bioorthogonal approach, ligand-directed chemistry recently emerged, but the slow kinetics limits its scope. Here we successfully overcome this obstacle using N-acyl-N-alkyl sulfonamide as a reactive group. Quantitative kinetic analyses reveal that ligand-directed N-acyl-N-alkyl sulfonamide chemistry allows for rapid modification of a lysine residue proximal to the ligand binding site of a target protein, with a rate constant of ~10(4) M(−1) s(−1), comparable to the fastest bioorthogonal chemistry. Despite some off-target reactions, this method can selectively label both intracellular and membrane-bound endogenous proteins. Moreover, the unique reactivity of N-acyl-N-alkyl sulfonamide enables the rational design of a lysine-targeted covalent inhibitor that shows durable suppression of the activity of Hsp90 in cancer cells. This work provides possibilities to extend the covalent inhibition approach that is currently being reassessed in drug discovery.
format Online
Article
Text
id pubmed-5951806
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-59518062018-05-16 Rapid labelling and covalent inhibition of intracellular native proteins using ligand-directed N-acyl-N-alkyl sulfonamide Tamura, Tomonori Ueda, Tsuyoshi Goto, Taiki Tsukidate, Taku Shapira, Yonatan Nishikawa, Yuki Fujisawa, Alma Hamachi, Itaru Nat Commun Article Selective modification of native proteins in live cells is one of the central challenges in recent chemical biology. As a unique bioorthogonal approach, ligand-directed chemistry recently emerged, but the slow kinetics limits its scope. Here we successfully overcome this obstacle using N-acyl-N-alkyl sulfonamide as a reactive group. Quantitative kinetic analyses reveal that ligand-directed N-acyl-N-alkyl sulfonamide chemistry allows for rapid modification of a lysine residue proximal to the ligand binding site of a target protein, with a rate constant of ~10(4) M(−1) s(−1), comparable to the fastest bioorthogonal chemistry. Despite some off-target reactions, this method can selectively label both intracellular and membrane-bound endogenous proteins. Moreover, the unique reactivity of N-acyl-N-alkyl sulfonamide enables the rational design of a lysine-targeted covalent inhibitor that shows durable suppression of the activity of Hsp90 in cancer cells. This work provides possibilities to extend the covalent inhibition approach that is currently being reassessed in drug discovery. Nature Publishing Group UK 2018-05-14 /pmc/articles/PMC5951806/ /pubmed/29760386 http://dx.doi.org/10.1038/s41467-018-04343-0 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Tamura, Tomonori
Ueda, Tsuyoshi
Goto, Taiki
Tsukidate, Taku
Shapira, Yonatan
Nishikawa, Yuki
Fujisawa, Alma
Hamachi, Itaru
Rapid labelling and covalent inhibition of intracellular native proteins using ligand-directed N-acyl-N-alkyl sulfonamide
title Rapid labelling and covalent inhibition of intracellular native proteins using ligand-directed N-acyl-N-alkyl sulfonamide
title_full Rapid labelling and covalent inhibition of intracellular native proteins using ligand-directed N-acyl-N-alkyl sulfonamide
title_fullStr Rapid labelling and covalent inhibition of intracellular native proteins using ligand-directed N-acyl-N-alkyl sulfonamide
title_full_unstemmed Rapid labelling and covalent inhibition of intracellular native proteins using ligand-directed N-acyl-N-alkyl sulfonamide
title_short Rapid labelling and covalent inhibition of intracellular native proteins using ligand-directed N-acyl-N-alkyl sulfonamide
title_sort rapid labelling and covalent inhibition of intracellular native proteins using ligand-directed n-acyl-n-alkyl sulfonamide
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5951806/
https://www.ncbi.nlm.nih.gov/pubmed/29760386
http://dx.doi.org/10.1038/s41467-018-04343-0
work_keys_str_mv AT tamuratomonori rapidlabellingandcovalentinhibitionofintracellularnativeproteinsusingliganddirectednacylnalkylsulfonamide
AT uedatsuyoshi rapidlabellingandcovalentinhibitionofintracellularnativeproteinsusingliganddirectednacylnalkylsulfonamide
AT gototaiki rapidlabellingandcovalentinhibitionofintracellularnativeproteinsusingliganddirectednacylnalkylsulfonamide
AT tsukidatetaku rapidlabellingandcovalentinhibitionofintracellularnativeproteinsusingliganddirectednacylnalkylsulfonamide
AT shapirayonatan rapidlabellingandcovalentinhibitionofintracellularnativeproteinsusingliganddirectednacylnalkylsulfonamide
AT nishikawayuki rapidlabellingandcovalentinhibitionofintracellularnativeproteinsusingliganddirectednacylnalkylsulfonamide
AT fujisawaalma rapidlabellingandcovalentinhibitionofintracellularnativeproteinsusingliganddirectednacylnalkylsulfonamide
AT hamachiitaru rapidlabellingandcovalentinhibitionofintracellularnativeproteinsusingliganddirectednacylnalkylsulfonamide