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Noncovalently Associated Peptides Observed during Liquid Chromatography-Mass Spectrometry and Their Effect on Cross-Link Analyses

[Image: see text] Cross-linking mass spectrometry draws structural information from covalently linked peptide pairs. When these links do not match to previous structural models, they may indicate changes in protein conformation. Unfortunately, such links can also be the result of experimental error...

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Autores principales: Giese, Sven H., Belsom, Adam, Sinn, Ludwig, Fischer, Lutz, Rappsilber, Juri
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2019
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6396951/
https://www.ncbi.nlm.nih.gov/pubmed/30649854
http://dx.doi.org/10.1021/acs.analchem.8b04037
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author Giese, Sven H.
Belsom, Adam
Sinn, Ludwig
Fischer, Lutz
Rappsilber, Juri
author_facet Giese, Sven H.
Belsom, Adam
Sinn, Ludwig
Fischer, Lutz
Rappsilber, Juri
author_sort Giese, Sven H.
collection PubMed
description [Image: see text] Cross-linking mass spectrometry draws structural information from covalently linked peptide pairs. When these links do not match to previous structural models, they may indicate changes in protein conformation. Unfortunately, such links can also be the result of experimental error or artifacts. Here, we describe the observation of noncovalently associated peptides during liquid chromatography-mass spectrometry analysis, which can easily be misidentified as cross-linked. Strikingly, they often mismatch to the protein structure. Noncovalently associated peptides presumably form during ionization and can be distinguished from cross-linked peptides by observing coelution of the corresponding linear peptides in MS1 spectra, as well as the presence of the individual (intact) peptide fragments in MS2 spectra. To suppress noncovalent peptide formations, increasingly disruptive ionization settings can be used, such as in-source fragmentation.
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spelling pubmed-63969512019-03-04 Noncovalently Associated Peptides Observed during Liquid Chromatography-Mass Spectrometry and Their Effect on Cross-Link Analyses Giese, Sven H. Belsom, Adam Sinn, Ludwig Fischer, Lutz Rappsilber, Juri Anal Chem [Image: see text] Cross-linking mass spectrometry draws structural information from covalently linked peptide pairs. When these links do not match to previous structural models, they may indicate changes in protein conformation. Unfortunately, such links can also be the result of experimental error or artifacts. Here, we describe the observation of noncovalently associated peptides during liquid chromatography-mass spectrometry analysis, which can easily be misidentified as cross-linked. Strikingly, they often mismatch to the protein structure. Noncovalently associated peptides presumably form during ionization and can be distinguished from cross-linked peptides by observing coelution of the corresponding linear peptides in MS1 spectra, as well as the presence of the individual (intact) peptide fragments in MS2 spectra. To suppress noncovalent peptide formations, increasingly disruptive ionization settings can be used, such as in-source fragmentation. American Chemical Society 2019-01-16 2019-02-19 /pmc/articles/PMC6396951/ /pubmed/30649854 http://dx.doi.org/10.1021/acs.analchem.8b04037 Text en Copyright © 2019 American Chemical Society This is an open access article published under a Creative Commons Attribution (CC-BY) License (http://pubs.acs.org/page/policy/authorchoice_ccby_termsofuse.html) , which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
spellingShingle Giese, Sven H.
Belsom, Adam
Sinn, Ludwig
Fischer, Lutz
Rappsilber, Juri
Noncovalently Associated Peptides Observed during Liquid Chromatography-Mass Spectrometry and Their Effect on Cross-Link Analyses
title Noncovalently Associated Peptides Observed during Liquid Chromatography-Mass Spectrometry and Their Effect on Cross-Link Analyses
title_full Noncovalently Associated Peptides Observed during Liquid Chromatography-Mass Spectrometry and Their Effect on Cross-Link Analyses
title_fullStr Noncovalently Associated Peptides Observed during Liquid Chromatography-Mass Spectrometry and Their Effect on Cross-Link Analyses
title_full_unstemmed Noncovalently Associated Peptides Observed during Liquid Chromatography-Mass Spectrometry and Their Effect on Cross-Link Analyses
title_short Noncovalently Associated Peptides Observed during Liquid Chromatography-Mass Spectrometry and Their Effect on Cross-Link Analyses
title_sort noncovalently associated peptides observed during liquid chromatography-mass spectrometry and their effect on cross-link analyses
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6396951/
https://www.ncbi.nlm.nih.gov/pubmed/30649854
http://dx.doi.org/10.1021/acs.analchem.8b04037
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