A proteomics approach to understanding protein ubiquitination View Full Text


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Article Info

DATE

2003-08

AUTHORS

Junmin Peng, Daniel Schwartz, Joshua E Elias, Carson C Thoreen, Dongmei Cheng, Gerald Marsischky, Jeroen Roelofs, Daniel Finley, Steven P Gygi

ABSTRACT

There is a growing need for techniques that can identify and characterize protein modifications on a large or global scale. We report here a proteomics approach to enrich, recover, and identify ubiquitin conjugates from Saccharomyces cerevisiae lysate. Ubiquitin conjugates from a strain expressing 6xHis-tagged ubiquitin were isolated, proteolyzed with trypsin and analyzed by multidimensional liquid chromatography coupled with tandem mass spectrometry (LC/LC-MS/MS) for amino acid sequence determination. We identified 1,075 proteins from the sample. In addition, we detected 110 precise ubiquitination sites present in 72 ubiquitin-protein conjugates. Finally, ubiquitin itself was found to be modified at seven lysine residues providing evidence for unexpected diversity in polyubiquitin chain topology in vivo. The methodology described here provides a general tool for the large-scale analysis and characterization of protein ubiquitination. More... »

PAGES

921-926

References to SciGraph publications

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  • Journal

    TITLE

    Nature Biotechnology

    ISSUE

    8

    VOLUME

    21

    Author Affiliations

    Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nbt849

    DOI

    http://dx.doi.org/10.1038/nbt849

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1015740794

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/12872131


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    JSON-LD is a popular format for linked data which is fully compatible with JSON.

    curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1038/nbt849'

    N-Triples is a line-based linked data format ideal for batch operations.

    curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1038/nbt849'

    Turtle is a human-readable linked data format.

    curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1038/nbt849'

    RDF/XML is a standard XML format for linked data.

    curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1038/nbt849'


     

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