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Photocatalytic proximity labelling of MCL-1 by a BH3 ligand
Ligand-directed protein labelling allows the introduction of diverse chemical functionalities onto proteins without the need for genetically encoded tags. Here we report a method for the rapid labelling of a protein using a ruthenium-bipyridyl (Ru(II)(bpy)(3))-modified peptide designed to mimic an i...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7610391/ https://www.ncbi.nlm.nih.gov/pubmed/33763603 http://dx.doi.org/10.1038/s42004-019-0235-z |
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author | Beard, Hester A. Hauser, Jacob R. Walko, Martin George, Rachel M. Wilson, Andrew J. Bon, Robin S. |
author_facet | Beard, Hester A. Hauser, Jacob R. Walko, Martin George, Rachel M. Wilson, Andrew J. Bon, Robin S. |
author_sort | Beard, Hester A. |
collection | PubMed |
description | Ligand-directed protein labelling allows the introduction of diverse chemical functionalities onto proteins without the need for genetically encoded tags. Here we report a method for the rapid labelling of a protein using a ruthenium-bipyridyl (Ru(II)(bpy)(3))-modified peptide designed to mimic an interacting BH3 ligand within a BCL-2 family protein-protein interactions. Using sub-stoichiometric quantities of (Ru(II)(bpy)(3))-modified NOXA-B and irradiation with visible light for 1 min, the anti-apoptotic protein MCL-1 can be photolabelled with a variety of functional tags. In contrast with previous reports on Ru(II)(bpy)(3)-mediated photolabelling, tandem mass spectrometry experiments reveal that the labelling site is a cysteine residue of MCL-1. MCL-1 can be labelled selectively in mixtures with other proteins, including the structurally related BCL-2 member, BCL-x(L). These results demonstrate that proximity-induced photolabelling is applicable to interfaces that mediate protein-protein interactions, and pave the way towards future use of ligand-directed proximity labelling for dynamic analysis of the interactome of BCL-2 family proteins. |
format | Online Article Text |
id | pubmed-7610391 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
record_format | MEDLINE/PubMed |
spelling | pubmed-76103912021-03-23 Photocatalytic proximity labelling of MCL-1 by a BH3 ligand Beard, Hester A. Hauser, Jacob R. Walko, Martin George, Rachel M. Wilson, Andrew J. Bon, Robin S. Commun Chem Article Ligand-directed protein labelling allows the introduction of diverse chemical functionalities onto proteins without the need for genetically encoded tags. Here we report a method for the rapid labelling of a protein using a ruthenium-bipyridyl (Ru(II)(bpy)(3))-modified peptide designed to mimic an interacting BH3 ligand within a BCL-2 family protein-protein interactions. Using sub-stoichiometric quantities of (Ru(II)(bpy)(3))-modified NOXA-B and irradiation with visible light for 1 min, the anti-apoptotic protein MCL-1 can be photolabelled with a variety of functional tags. In contrast with previous reports on Ru(II)(bpy)(3)-mediated photolabelling, tandem mass spectrometry experiments reveal that the labelling site is a cysteine residue of MCL-1. MCL-1 can be labelled selectively in mixtures with other proteins, including the structurally related BCL-2 member, BCL-x(L). These results demonstrate that proximity-induced photolabelling is applicable to interfaces that mediate protein-protein interactions, and pave the way towards future use of ligand-directed proximity labelling for dynamic analysis of the interactome of BCL-2 family proteins. 2019-11-21 /pmc/articles/PMC7610391/ /pubmed/33763603 http://dx.doi.org/10.1038/s42004-019-0235-z Text en https://creativecommons.org/licenses/by/4.0/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 https://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Beard, Hester A. Hauser, Jacob R. Walko, Martin George, Rachel M. Wilson, Andrew J. Bon, Robin S. Photocatalytic proximity labelling of MCL-1 by a BH3 ligand |
title | Photocatalytic proximity labelling of MCL-1 by a BH3 ligand |
title_full | Photocatalytic proximity labelling of MCL-1 by a BH3 ligand |
title_fullStr | Photocatalytic proximity labelling of MCL-1 by a BH3 ligand |
title_full_unstemmed | Photocatalytic proximity labelling of MCL-1 by a BH3 ligand |
title_short | Photocatalytic proximity labelling of MCL-1 by a BH3 ligand |
title_sort | photocatalytic proximity labelling of mcl-1 by a bh3 ligand |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7610391/ https://www.ncbi.nlm.nih.gov/pubmed/33763603 http://dx.doi.org/10.1038/s42004-019-0235-z |
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