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Cross-Linking Electrochemical Mass Spectrometry for Probing Protein Three-Dimensional Structures

[Image: see text] Chemical cross-linking combined with mass spectrometry (MS) is powerful to provide protein three-dimensional structure information but difficulties in identifying cross-linked peptides and elucidating their structures limit its usefulness. To tackle these challenges, this study pre...

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Autores principales: Zheng, Qiuling, Zhang, Hao, Tong, Lingying, Wu, Shiyong, Chen, Hao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2014
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4165463/
https://www.ncbi.nlm.nih.gov/pubmed/25141260
http://dx.doi.org/10.1021/ac501526n
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author Zheng, Qiuling
Zhang, Hao
Tong, Lingying
Wu, Shiyong
Chen, Hao
author_facet Zheng, Qiuling
Zhang, Hao
Tong, Lingying
Wu, Shiyong
Chen, Hao
author_sort Zheng, Qiuling
collection PubMed
description [Image: see text] Chemical cross-linking combined with mass spectrometry (MS) is powerful to provide protein three-dimensional structure information but difficulties in identifying cross-linked peptides and elucidating their structures limit its usefulness. To tackle these challenges, this study presents a novel cross-linking MS in conjunction with electrochemistry using disulfide-bond-containing dithiobis[succinimidyl propionate] (DSP) as the cross-linker. In our approach, electrolysis of DSP-bridged protein/peptide products, as online monitored by desorption electrospray ionization mass spectrometry is highly informative. First, as disulfide bonds are electrochemically reducible, the cross-links are subject to pronounced intensity decrease upon electrolytic reduction, suggesting a new way to identify cross-links. Also, mass shift before and after electrolysis suggests the linkage pattern of cross-links. Electrochemical reduction removes disulfide bond constraints, possibly increasing sequence coverage for tandem MS analysis and yielding linear peptides whose structures are more easily determined than their cross-linked precursor peptides. Furthermore, this cross-linking electrochemical MS method is rapid, due to the fast nature of electrochemical conversion (much faster than traditional chemical reduction) and no need for chromatographic separation, which would be of high value for structural proteomics research.
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spelling pubmed-41654632015-08-20 Cross-Linking Electrochemical Mass Spectrometry for Probing Protein Three-Dimensional Structures Zheng, Qiuling Zhang, Hao Tong, Lingying Wu, Shiyong Chen, Hao Anal Chem [Image: see text] Chemical cross-linking combined with mass spectrometry (MS) is powerful to provide protein three-dimensional structure information but difficulties in identifying cross-linked peptides and elucidating their structures limit its usefulness. To tackle these challenges, this study presents a novel cross-linking MS in conjunction with electrochemistry using disulfide-bond-containing dithiobis[succinimidyl propionate] (DSP) as the cross-linker. In our approach, electrolysis of DSP-bridged protein/peptide products, as online monitored by desorption electrospray ionization mass spectrometry is highly informative. First, as disulfide bonds are electrochemically reducible, the cross-links are subject to pronounced intensity decrease upon electrolytic reduction, suggesting a new way to identify cross-links. Also, mass shift before and after electrolysis suggests the linkage pattern of cross-links. Electrochemical reduction removes disulfide bond constraints, possibly increasing sequence coverage for tandem MS analysis and yielding linear peptides whose structures are more easily determined than their cross-linked precursor peptides. Furthermore, this cross-linking electrochemical MS method is rapid, due to the fast nature of electrochemical conversion (much faster than traditional chemical reduction) and no need for chromatographic separation, which would be of high value for structural proteomics research. American Chemical Society 2014-08-20 2014-09-16 /pmc/articles/PMC4165463/ /pubmed/25141260 http://dx.doi.org/10.1021/ac501526n Text en Copyright © 2014 American Chemical Society Terms of Use (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html)
spellingShingle Zheng, Qiuling
Zhang, Hao
Tong, Lingying
Wu, Shiyong
Chen, Hao
Cross-Linking Electrochemical Mass Spectrometry for Probing Protein Three-Dimensional Structures
title Cross-Linking Electrochemical Mass Spectrometry for Probing Protein Three-Dimensional Structures
title_full Cross-Linking Electrochemical Mass Spectrometry for Probing Protein Three-Dimensional Structures
title_fullStr Cross-Linking Electrochemical Mass Spectrometry for Probing Protein Three-Dimensional Structures
title_full_unstemmed Cross-Linking Electrochemical Mass Spectrometry for Probing Protein Three-Dimensional Structures
title_short Cross-Linking Electrochemical Mass Spectrometry for Probing Protein Three-Dimensional Structures
title_sort cross-linking electrochemical mass spectrometry for probing protein three-dimensional structures
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4165463/
https://www.ncbi.nlm.nih.gov/pubmed/25141260
http://dx.doi.org/10.1021/ac501526n
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