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An integrated workflow for crosslinking mass spectrometry.

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.

SUBMITTER: Mendes ML

PROVIDER: S-EPMC6753376 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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An integrated workflow for crosslinking mass spectrometry.

Mendes Marta L ML Fischer Lutz L Chen Zhuo A ZA Barbon Marta M O'Reilly Francis J FJ Giese Sven H SH Bohlke-Schneider Michael M Belsom Adam A Dau Therese T Combe Colin W CW Graham Martin M Eisele Markus R MR Baumeister Wolfgang W Speck Christian C Rappsilber Juri J

Molecular systems biology 20190901 9

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 ove ...[more]

PMID: 31556486

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