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  An integrated workflow for crosslinking mass spectrometry

Mendes, M. L., Fischer, L., Chen, Z. A., Barbon, M., O'Reilly, F. J., Giese, S. H., et al. (2019). An integrated workflow for crosslinking mass spectrometry. MOLECULAR SYSTEMS BIOLOGY, 15(9): e8994. doi:10.15252/msb.20198994.

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Mendes_et_al-2019-Molecular_Systems_Biology.pdf (Any fulltext), 3MB
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 Creators:
Mendes, Marta L.1, Author
Fischer, Lutz1, Author
Chen, Zhuo A.1, Author
Barbon, Marta1, Author
O'Reilly, Francis J.1, Author
Giese, Sven H.1, Author
Bohlke-Schneider, Michael1, Author
Belsom, Adam1, Author
Dau, Therese1, Author
Combe, Colin W.1, Author
Graham, Martin1, Author
Eisele, Markus R.2, Author              
Baumeister, Wolfgang2, Author              
Speck, Christian1, Author
Rappsilber, Juri1, Author
Affiliations:
1external, ou_persistent22              
2Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              

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Free keywords: PROTEIN INTERACTIONS; STRUCTURAL BASIS; C5 CONVERTASE; DIGESTION; PROTEOMICS; MCM2-7; DNA; IDENTIFICATION; FRAGMENTATION; ARCHITECTUREBiochemistry & Molecular Biology; crosslinking mass spectrometry; protein-protein interactions; proteomics; software; structural biology;
 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.

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Language(s): eng - English
 Dates: 2019
 Publication Status: Published online
 Pages: 13
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000488972600003
DOI: 10.15252/msb.20198994
 Degree: -

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Title: MOLECULAR SYSTEMS BIOLOGY
Source Genre: Journal
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Publ. Info: 111 RIVER ST, HOBOKEN 07030-5774, NJ USA : WILEY
Pages: - Volume / Issue: 15 (9) Sequence Number: e8994 Start / End Page: - Identifier: ISSN: 1744-4292