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pH Dependence of Succinimide-Ester-Based Protein Cross-Linking for Structural Mass Spectrometry Applications
[Image: see text] Within the research field of cross-linking mass spectrometry (XL-MS), the most commonly used cross-linking reagents are succinimide-ester-based (e.g., disuccinimidyl suberate (DSS)). These reagents primarily cross-link lysine side chains. So far, they have predominantly been used t...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9838815/ https://www.ncbi.nlm.nih.gov/pubmed/36785722 http://dx.doi.org/10.1021/acsmeasuresciau.1c00032 |
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author | Trabjerg, Esben Keller, Arend Leitner, Alexander |
author_facet | Trabjerg, Esben Keller, Arend Leitner, Alexander |
author_sort | Trabjerg, Esben |
collection | PubMed |
description | [Image: see text] Within the research field of cross-linking mass spectrometry (XL-MS), the most commonly used cross-linking reagents are succinimide-ester-based (e.g., disuccinimidyl suberate (DSS)). These reagents primarily cross-link lysine side chains. So far, they have predominantly been used to investigate protein structures at neutral to slightly basic pH (7.0–8.5) to ensure the reactivity of the primary amine of the lysine side chain. However, disease-related molecular processes are not limited to such pH ranges; e.g., some important biological pathways are active in acidic intracellular compartments. The applicability of lysine-reactive cross-linking reagents to low-pH conditions remains unclear. Here, we cross-linked a mixture of eight model proteins at eight different pH conditions (pH 4.0–7.5) to investigate the pH dependency of DSS. DSS was able to cross-link proteins even at pH 4.0, but a clear decrease in the cross-linking efficiency was observed when the pH was lowered. Nevertheless, at pH 5.0, approximately half of the number of cross-links observed at pH 7.5 could still be identified. These findings highlight the ability of succinimide-based cross-linking reagents to be useful in probing the structure of proteins in a slightly acidic environment. |
format | Online Article Text |
id | pubmed-9838815 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-98388152023-02-10 pH Dependence of Succinimide-Ester-Based Protein Cross-Linking for Structural Mass Spectrometry Applications Trabjerg, Esben Keller, Arend Leitner, Alexander ACS Meas Sci Au [Image: see text] Within the research field of cross-linking mass spectrometry (XL-MS), the most commonly used cross-linking reagents are succinimide-ester-based (e.g., disuccinimidyl suberate (DSS)). These reagents primarily cross-link lysine side chains. So far, they have predominantly been used to investigate protein structures at neutral to slightly basic pH (7.0–8.5) to ensure the reactivity of the primary amine of the lysine side chain. However, disease-related molecular processes are not limited to such pH ranges; e.g., some important biological pathways are active in acidic intracellular compartments. The applicability of lysine-reactive cross-linking reagents to low-pH conditions remains unclear. Here, we cross-linked a mixture of eight model proteins at eight different pH conditions (pH 4.0–7.5) to investigate the pH dependency of DSS. DSS was able to cross-link proteins even at pH 4.0, but a clear decrease in the cross-linking efficiency was observed when the pH was lowered. Nevertheless, at pH 5.0, approximately half of the number of cross-links observed at pH 7.5 could still be identified. These findings highlight the ability of succinimide-based cross-linking reagents to be useful in probing the structure of proteins in a slightly acidic environment. American Chemical Society 2021-11-11 /pmc/articles/PMC9838815/ /pubmed/36785722 http://dx.doi.org/10.1021/acsmeasuresciau.1c00032 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Trabjerg, Esben Keller, Arend Leitner, Alexander pH Dependence of Succinimide-Ester-Based Protein Cross-Linking for Structural Mass Spectrometry Applications |
title | pH Dependence of Succinimide-Ester-Based Protein Cross-Linking
for Structural Mass Spectrometry Applications |
title_full | pH Dependence of Succinimide-Ester-Based Protein Cross-Linking
for Structural Mass Spectrometry Applications |
title_fullStr | pH Dependence of Succinimide-Ester-Based Protein Cross-Linking
for Structural Mass Spectrometry Applications |
title_full_unstemmed | pH Dependence of Succinimide-Ester-Based Protein Cross-Linking
for Structural Mass Spectrometry Applications |
title_short | pH Dependence of Succinimide-Ester-Based Protein Cross-Linking
for Structural Mass Spectrometry Applications |
title_sort | ph dependence of succinimide-ester-based protein cross-linking
for structural mass spectrometry applications |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9838815/ https://www.ncbi.nlm.nih.gov/pubmed/36785722 http://dx.doi.org/10.1021/acsmeasuresciau.1c00032 |
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