An integrated workflow for crosslinking mass spectrometry.
crosslinking mass spectrometry
protein-protein interactions
proteomics
software
structural biology
Journal
Molecular systems biology
ISSN: 1744-4292
Titre abrégé: Mol Syst Biol
Pays: England
ID NLM: 101235389
Informations de publication
Date de publication:
09 2019
09 2019
Historique:
received:
09
05
2019
revised:
19
08
2019
accepted:
21
08
2019
entrez:
27
9
2019
pubmed:
27
9
2019
medline:
21
5
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31556486
doi: 10.15252/msb.20198994
pmc: PMC6753376
doi:
Substances chimiques
Peptide Fragments
0
Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e8994Subventions
Organisme : Wellcome Trust
ID : 203149
Pays : United Kingdom
Organisme : UK Research and Innovation | Biotechnology and Biological Sciences Research Council (BBSRC)
ID : BB/N000323/1
Pays : International
Organisme : Wellcome Trust
ID : 108504
Pays : United Kingdom
Organisme : UK Research and Innovation | Medical Research Council (MRC)
ID : MC_U120085811
Pays : International
Organisme : Wellcome Trust
ID : 103139/Z/13/Z
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 103139
Pays : United Kingdom
Organisme : Wellcome Trust (WT)
ID : 107903/Z/15/Z
Pays : International
Organisme : Medical Research Council
ID : MC_U120085811
Pays : United Kingdom
Informations de copyright
© 2019 The Authors. Published under the terms of the CC BY 4.0 license.
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