Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Trabjerg, Esben, Keller, Arend, Leitner, Alexander
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2021
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
_version_ 1784869357555286016
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
work_keys_str_mv AT trabjergesben phdependenceofsuccinimideesterbasedproteincrosslinkingforstructuralmassspectrometryapplications
AT kellerarend phdependenceofsuccinimideesterbasedproteincrosslinkingforstructuralmassspectrometryapplications
AT leitneralexander phdependenceofsuccinimideesterbasedproteincrosslinkingforstructuralmassspectrometryapplications