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  • Brief Communication
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Identification of cross-linked peptides from large sequence databases

A Corrigendum to this article was published on 01 August 2008

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NOTE: In the version of this Brief Communication initially published, an author name (Lukas Mueller) was incorrect. The correct author name is Lukas N Mueller. The error has been corrected in the HTML and PDF versions of the article.

We describe a method to identify cross-linked peptides from complex samples and large protein sequence databases by combining isotopically tagged cross-linkers, chromatographic enrichment, targeted proteomics and a new search engine called xQuest. This software reduces the search space by an upstream candidate-peptide search before the recombination step. We showed that xQuest can identify cross-linked peptides from a total Escherichia coli lysate with an unrestricted database search.

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Figure 1: Workflow of cross-link search with xQuest in the ion-tag mode.
Figure 2: Linear discriminant analysis separates true positive MS2 assignments to standard protein cross-links from false positive hits in a large peptide-interaction database.
Figure 3: Inter-protein and intra-protein cross-links were identified from a total E. coli lysate, searched against the total E. coli protein database.

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Change history

  • 27 June 2008

    In the version of this Brief Communication initially published, an author name (Lukas Mueller) was incorrect. The correct author name is Lukas N Mueller. The error has been corrected in the HTML and PDF versions of the article.


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O.R. was supported by fellowships of the Deutsche Forschungsgemeinschaft and the Roche Research Foundation; M.B. was supported by a long-term fellowship of the European Molecular Biology Organization and a Marie Curie fellowship of the European Commission. A.S. was supported in part by a grant from F. Hoffmann–La Roche Ltd provided to the Competence Center for Systems Physiology and Metabolic Disease. The work was supported in part with Federal funds from the US National Heart, Lung and Blood Institute, National Institutes of Health (contract N01-HV-28179) and with funds from ETH Zurich.

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Authors and Affiliations



O.R. was responsible for most of the software design and programming, conducted most of the experiments and most of the data evaluation; J.S. synthesized the cross-linker and contributed to software design; T.W. contributed to scoring, data evaluation and part of experiments; L.N.M. analyzed and aligned MS1 data; M.B. performed part of experiments and evaluated data; A.S. designed mass spectrometric methods for analyzing cross-linked samples; M.M. contributed to scoring; R.A. coordinated the project, contributed ideas and concept.

Corresponding author

Correspondence to Ruedi Aebersold.

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Supplementary Figures 1–5, Supplementary Tables 1–2, Supplementary Results, Supplementary Discussion, Supplementary Methods (PDF 1982 kb)

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Rinner, O., Seebacher, J., Walzthoeni, T. et al. Identification of cross-linked peptides from large sequence databases. Nat Methods 5, 315–318 (2008).

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