Denaturing mass photometry for rapid optimization of chemical protein-protein cross-linking reactions
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2024 |
| Outros Autores: | , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
| Texto Completo: | http://hdl.handle.net/10362/172803 |
Resumo: | Funding Information: This work was supported by the CNRS, the University of Strasbourg, Lorraine University, the \u201CAgence National de la Recherche\u201D (ANR-16-CE11-0032-04 snoRNP assembly; ANR-23-CE44-0035 NAProt-XLMS) and the French Proteomics Infrastructure (ProFI; ANR-10-INBS-08-03). H.G.F. acknowledges the French Ministry for Education and Research for funding of his PhD. This work was funded by Funda\u00E7\u00E3o para a Ci\u00EAncia e Tecnologia/Minist\u00E9rio da Ci\u00EAncia, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through national funds to iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). Publisher Copyright: © The Author(s) 2024. |
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Denaturing mass photometry for rapid optimization of chemical protein-protein cross-linking reactionsChemistry(all)Biochemistry, Genetics and Molecular Biology(all)Physics and Astronomy(all)Funding Information: This work was supported by the CNRS, the University of Strasbourg, Lorraine University, the \u201CAgence National de la Recherche\u201D (ANR-16-CE11-0032-04 snoRNP assembly; ANR-23-CE44-0035 NAProt-XLMS) and the French Proteomics Infrastructure (ProFI; ANR-10-INBS-08-03). H.G.F. acknowledges the French Ministry for Education and Research for funding of his PhD. This work was funded by Funda\u00E7\u00E3o para a Ci\u00EAncia e Tecnologia/Minist\u00E9rio da Ci\u00EAncia, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through national funds to iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). Publisher Copyright: © The Author(s) 2024.Chemical cross-linking reactions (XL) are an important strategy for studying protein-protein interactions (PPIs), including low abundant sub-complexes, in structural biology. However, choosing XL reagents and conditions is laborious and mostly limited to analysis of protein assemblies that can be resolved using SDS-PAGE. To overcome these limitations, we develop here a denaturing mass photometry (dMP) method for fast, reliable and user-friendly optimization and monitoring of chemical XL reactions. The dMP is a robust 2-step protocol that ensures 95% of irreversible denaturation within only 5 min. We show that dMP provides accurate mass identification across a broad mass range (30 kDa–5 MDa) along with direct label-free relative quantification of all coexisting XL species (sub-complexes and aggregates). We compare dMP with SDS-PAGE and observe that, unlike the benchmark, dMP is time-efficient (3 min/triplicate), requires significantly less material (20–100×) and affords single molecule sensitivity. To illustrate its utility for routine structural biology applications, we show that dMP affords screening of 20 XL conditions in 1 h, accurately identifying and quantifying all coexisting species. Taken together, we anticipate that dMP will have an impact on ability to structurally characterize more PPIs and macromolecular assemblies, expected final complexes but also sub-complexes that form en route.Instituto de Tecnologia Química e Biológica António Xavier (ITQB)RUNGizardin-Fredon, HugoSanto, Paulo E.Chagot, Marie EveCharpentier, BrunoBandeiras, Tiago M.Manival, XavierHernandez-Alba, OscarCianférani, Sarah2024-10-01T22:22:41Z2024-04-252024-04-25T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/172803eng2041-1723PURE: 90361378https://doi.org/10.1038/s41467-024-47732-4info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-10-07T01:39:53Zoai:run.unl.pt:10362/172803Portal AgregadorONGhttps://www.rcaap.pt/oai/openairemluisa.alvim@gmail.comopendoar:71602024-10-07T01:39:53Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
| dc.title.none.fl_str_mv |
Denaturing mass photometry for rapid optimization of chemical protein-protein cross-linking reactions |
| title |
Denaturing mass photometry for rapid optimization of chemical protein-protein cross-linking reactions |
| spellingShingle |
Denaturing mass photometry for rapid optimization of chemical protein-protein cross-linking reactions Gizardin-Fredon, Hugo Chemistry(all) Biochemistry, Genetics and Molecular Biology(all) Physics and Astronomy(all) |
| title_short |
Denaturing mass photometry for rapid optimization of chemical protein-protein cross-linking reactions |
| title_full |
Denaturing mass photometry for rapid optimization of chemical protein-protein cross-linking reactions |
| title_fullStr |
Denaturing mass photometry for rapid optimization of chemical protein-protein cross-linking reactions |
| title_full_unstemmed |
Denaturing mass photometry for rapid optimization of chemical protein-protein cross-linking reactions |
| title_sort |
Denaturing mass photometry for rapid optimization of chemical protein-protein cross-linking reactions |
| author |
Gizardin-Fredon, Hugo |
| author_facet |
Gizardin-Fredon, Hugo Santo, Paulo E. Chagot, Marie Eve Charpentier, Bruno Bandeiras, Tiago M. Manival, Xavier Hernandez-Alba, Oscar Cianférani, Sarah |
| author_role |
author |
| author2 |
Santo, Paulo E. Chagot, Marie Eve Charpentier, Bruno Bandeiras, Tiago M. Manival, Xavier Hernandez-Alba, Oscar Cianférani, Sarah |
| author2_role |
author author author author author author author |
| dc.contributor.none.fl_str_mv |
Instituto de Tecnologia Química e Biológica António Xavier (ITQB) RUN |
| dc.contributor.author.fl_str_mv |
Gizardin-Fredon, Hugo Santo, Paulo E. Chagot, Marie Eve Charpentier, Bruno Bandeiras, Tiago M. Manival, Xavier Hernandez-Alba, Oscar Cianférani, Sarah |
| dc.subject.por.fl_str_mv |
Chemistry(all) Biochemistry, Genetics and Molecular Biology(all) Physics and Astronomy(all) |
| topic |
Chemistry(all) Biochemistry, Genetics and Molecular Biology(all) Physics and Astronomy(all) |
| description |
Funding Information: This work was supported by the CNRS, the University of Strasbourg, Lorraine University, the \u201CAgence National de la Recherche\u201D (ANR-16-CE11-0032-04 snoRNP assembly; ANR-23-CE44-0035 NAProt-XLMS) and the French Proteomics Infrastructure (ProFI; ANR-10-INBS-08-03). H.G.F. acknowledges the French Ministry for Education and Research for funding of his PhD. This work was funded by Funda\u00E7\u00E3o para a Ci\u00EAncia e Tecnologia/Minist\u00E9rio da Ci\u00EAncia, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through national funds to iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). Publisher Copyright: © The Author(s) 2024. |
| publishDate |
2024 |
| dc.date.none.fl_str_mv |
2024-10-01T22:22:41Z 2024-04-25 2024-04-25T00:00:00Z |
| dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/172803 |
| url |
http://hdl.handle.net/10362/172803 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
2041-1723 PURE: 90361378 https://doi.org/10.1038/s41467-024-47732-4 |
| dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
| eu_rights_str_mv |
openAccess |
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application/pdf |
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reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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mluisa.alvim@gmail.com |
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1817548564179451904 |