ArticlePloS one2012
Detection of crosslinks within and between proteins by LC-MALDI-TOFTOF and the software FINDX to reduce the MSMS-data to acquire for validation.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Article
- Applications of MALDI-MS/MS-Based Proteomics in Biomedical Research.Molecules (Basel, Switzerland) · 2022Review
- Structural dynamics of the E6AP/UBE3A-E6-p53 enzyme-substrate complex.Nature communications · 2018Article
- Structural model of dodecameric heat-shock protein Hsp21: Flexible N-terminal arms interact with client proteins while C-terminal tails maintain the dodecamer and chaperone activity.The Journal of biological chemistry · 2017Article
- High Sensitivity Crosslink Detection Coupled With Integrative Structure Modeling in the Mass Spec Studio.Molecular & cellular proteomics : MCP · 2016Article
- The Structure of the Complex between Yeast Frataxin and Ferrochelatase: CHARACTERIZATION AND PRE-STEADY STATE REACTION OF FERROUS IRON DELIVERY AND HEME SYNTHESIS.The Journal of biological chemistry · 2016Article
- Chemical cross-linking and native mass spectrometry: A fruitful combination for structural biology.Protein science : a publication of the Protein Society · 2015Review
- Article
- Technical note: proteomic approaches to fundamental questions about neutrophil biology.Journal of leukocyte biology · 2013Review
- Cross-linking and mass spectrometry methodologies to facilitate structural biology: finding a path through the maze.Journal of structural and functional genomics · 2013Review
- Semi-automated liquid chromatography-mass spectrometric imaging platform for enhanced detection and improved data analysis of complex peptides.Journal of chromatography. A · 2013Article
- Mixed-isotope labeling with LC-IMS-MS for characterization of protein-protein interactions by chemical cross-linking.Journal of the American Society for Mass Spectrometry · 2013Article
- Probing the transient interaction between the small heat-shock protein Hsp21 and a model substrate protein using crosslinking mass spectrometry.Cell stress & chaperones · 2013Article
Corrections and comments
- Erratum issued
Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Lysine-specific chemical crosslinking in combination with mass spectrometry is emerging as a tool for the structural characterization of protein complexes and protein-protein interactions. After tryptic digestion of crosslinked proteins there are thousands of peptides amenable to MSMS, of which only very few are crosslinked peptides of interest. Here we describe how the advantage offered by off-line LC-MALDI-TOF/TOF mass spectrometry is exploited in a two-step workflow to focus the MSMS-acquisition on crosslinks mainly. In a first step, MS-data are acquired and all the peak list files from the LC-separated fractions are merged by the FINDX software and screened for presence of crosslinks which are recognized as isotope-labeled doublet peaks. Information on the isotope doublet peak mass and intensity can be used as search constraints to reduce the number of false positives that match randomly to the observed peak masses. Based on the MS-data a precursor ion inclusion list is generated and used in a second step, where a restricted number of MSMS-spectra are acquired for crosslink validation. The decoupling of MS and MSMS and the peptide sorting with FINDX based on MS-data has the advantage that MSMS can be restricted to and focused on crosslinks of Type 2, which are of highest biological interest but often lowest in abundance. The LC-MALDI TOF/TOF workflow here described is applicable to protein multisubunit complexes and using (14)N/(15)N mixed isotope strategy for the detection of inter-protein crosslinks within protein oligomers.
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