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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American
Chemical
Society
2014
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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. |
format | Online Article Text |
id | pubmed-4165463 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | American
Chemical
Society |
record_format | MEDLINE/PubMed |
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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