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An integrated workflow for crosslinking mass spectrometry

We present a concise workflow to enhance the mass spectrometric detection of crosslinked peptides by introducing sequential digestion and the crosslink identification software xiSEARCH. Sequential digestion enhances peptide detection by selective shortening of long tryptic peptides. We demonstrate o...

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Detalles Bibliográficos
Autores principales: Mendes, Marta L, Fischer, Lutz, Chen, Zhuo A, Barbon, Marta, O'Reilly, Francis J, Giese, Sven H, Bohlke‐Schneider, Michael, Belsom, Adam, Dau, Therese, Combe, Colin W, Graham, Martin, Eisele, Markus R, Baumeister, Wolfgang, Speck, Christian, Rappsilber, Juri
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6753376/
https://www.ncbi.nlm.nih.gov/pubmed/31556486
http://dx.doi.org/10.15252/msb.20198994
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author Mendes, Marta L
Fischer, Lutz
Chen, Zhuo A
Barbon, Marta
O'Reilly, Francis J
Giese, Sven H
Bohlke‐Schneider, Michael
Belsom, Adam
Dau, Therese
Combe, Colin W
Graham, Martin
Eisele, Markus R
Baumeister, Wolfgang
Speck, Christian
Rappsilber, Juri
author_facet Mendes, Marta L
Fischer, Lutz
Chen, Zhuo A
Barbon, Marta
O'Reilly, Francis J
Giese, Sven H
Bohlke‐Schneider, Michael
Belsom, Adam
Dau, Therese
Combe, Colin W
Graham, Martin
Eisele, Markus R
Baumeister, Wolfgang
Speck, Christian
Rappsilber, Juri
author_sort Mendes, Marta L
collection PubMed
description We present a concise workflow to enhance the mass spectrometric detection of crosslinked peptides by introducing sequential digestion and the crosslink identification software xiSEARCH. Sequential digestion enhances peptide detection by selective shortening of long tryptic peptides. We demonstrate our simple 12‐fraction protocol for crosslinked multi‐protein complexes and cell lysates, quantitative analysis, and high‐density crosslinking, without requiring specific crosslinker features. This overall approach reveals dynamic protein–protein interaction sites, which are accessible, have fundamental functional relevance and are therefore ideally suited for the development of small molecule inhibitors.
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spelling pubmed-67533762019-09-23 An integrated workflow for crosslinking mass spectrometry Mendes, Marta L Fischer, Lutz Chen, Zhuo A Barbon, Marta O'Reilly, Francis J Giese, Sven H Bohlke‐Schneider, Michael Belsom, Adam Dau, Therese Combe, Colin W Graham, Martin Eisele, Markus R Baumeister, Wolfgang Speck, Christian Rappsilber, Juri Mol Syst Biol Methods We present a concise workflow to enhance the mass spectrometric detection of crosslinked peptides by introducing sequential digestion and the crosslink identification software xiSEARCH. Sequential digestion enhances peptide detection by selective shortening of long tryptic peptides. We demonstrate our simple 12‐fraction protocol for crosslinked multi‐protein complexes and cell lysates, quantitative analysis, and high‐density crosslinking, without requiring specific crosslinker features. This overall approach reveals dynamic protein–protein interaction sites, which are accessible, have fundamental functional relevance and are therefore ideally suited for the development of small molecule inhibitors. John Wiley and Sons Inc. 2019-09-20 /pmc/articles/PMC6753376/ /pubmed/31556486 http://dx.doi.org/10.15252/msb.20198994 Text en © 2019 The Authors. Published under the terms of the CC BY 4.0 license This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods
Mendes, Marta L
Fischer, Lutz
Chen, Zhuo A
Barbon, Marta
O'Reilly, Francis J
Giese, Sven H
Bohlke‐Schneider, Michael
Belsom, Adam
Dau, Therese
Combe, Colin W
Graham, Martin
Eisele, Markus R
Baumeister, Wolfgang
Speck, Christian
Rappsilber, Juri
An integrated workflow for crosslinking mass spectrometry
title An integrated workflow for crosslinking mass spectrometry
title_full An integrated workflow for crosslinking mass spectrometry
title_fullStr An integrated workflow for crosslinking mass spectrometry
title_full_unstemmed An integrated workflow for crosslinking mass spectrometry
title_short An integrated workflow for crosslinking mass spectrometry
title_sort integrated workflow for crosslinking mass spectrometry
topic Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6753376/
https://www.ncbi.nlm.nih.gov/pubmed/31556486
http://dx.doi.org/10.15252/msb.20198994
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