Open AccessAn integrated workflow for crosslinking mass spectrometry
Author(s) -
Mendes Marta L,
Fischer Lutz,
Chen Zhuo A,
Barbon Marta,
O'Reilly Francis J,
Giese Sven H,
BohlkeSchneider Michael,
Belsom Adam,
Dau Therese,
Combe Colin W,
Graham Martin,
Eisele Markus R,
Baumeister Wolfgang,
Speck Christian,
Rappsilber Juri
Publication year - 2019
Publication title -
molecular systems biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 8.523
H-Index - 148
ISSN - 1744-4292
DOI - 10.15252/msb.20198994
Subject(s) - workflow , mass spectrometry , peptide , biology , computational biology , digestion (alchemy) , combinatorial chemistry , biochemistry , chromatography , computer science , chemistry , database
We present a concise workflow to enhance the mass spectrometric detection of crosslinked peptides by introducing sequential digestion and the crosslink identification software xi SEARCH . 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.