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An integrated workflow for crosslinking mass spectrometry

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Title: An integrated workflow for crosslinking mass spectrometry
Authors: Mendes, ML
Fischer, L
Chen, ZA
Barbon, M
O'Reilly, FJ
Giese, SH
Bohlke-Schneider, M
Belsom, A
Dau, T
Combe, CW
Graham, M
Eisele, MR
Baumeister, W
Speck, C
Rappsilber, J
Item Type: Journal Article
Abstract: 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.
Issue Date: 1-Sep-2019
Date of Acceptance: 21-Aug-2019
URI: http://hdl.handle.net/10044/1/74010
DOI: https://dx.doi.org/10.15252/msb.20198994
ISSN: 1744-4292
Publisher: European Molecular Biology Organization
Journal / Book Title: Molecular Systems Biology
Volume: 15
Issue: 9
Copyright Statement: © 2019The Authors. Published under the terms of the CC- BY 4.0 license. This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Sponsor/Funder: Wellcome Trust
Biotechnology and Biological Sciences Research Council (BBSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Funder's Grant Number: 107903/Z/15/Z
Keywords: crosslinking mass spectrometry
protein-protein interactions
structural biology
0699 Other Biological Sciences
0601 Biochemistry and Cell Biology
Publication Status: Published
Open Access location: https://www.embopress.org/doi/full/10.15252/msb.20198994
Article Number: ARTN e8994
Appears in Collections:Institute of Clinical Sciences