Proteins interacting with Saccharomyces cerevisiae type 1 protein phosphatase catalytic subunit identified by single-step affinity purification and mass spectrometry

Edmund P. Walsh, Douglas Lamont, Kenneth A. Beattie, Michael J. R. Stark

    Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)

    1 Citation (Scopus)

    Abstract

    The catalytic subunit of type 1 protein phosphatase (PP1C) interacts with a large number of polypeptides in eukaryotic cells from yeast to man and these regulatory subunits can both modulate the activity of PP1C and target it to different subcellular locations. Thus, PP1 is really a family of protein phosphatases that share a common catalytic subunit, and identifying and characterizing the PP1-associated proteins is therefore critical to understanding the cellular roles of PP1 and its ability to dephosphorylate specific substrates. Here we describe methods for affinity isolation of PP1C-containing protein complexes in the yeast Saccharomyces cerevisiae and the identification of the associated polypeptides by mass spectrometry. The basic method we describe could be easily adapted to study PP1C-associated proteins in other lower or higher eukaryotes and for characterizing the protein-protein interactions of other protein phosphatases in yeast.

    Original languageEnglish
    Title of host publicationProtein phosphatase protocols
    EditorsGreg Moorhead
    PublisherHumana Press
    Pages235-246
    Number of pages12
    ISBN (Electronic)9781597452670
    ISBN (Print)9781588297112
    DOIs
    Publication statusPublished - 2007

    Publication series

    NameMethods in Molecular Biology
    Volume365
    ISSN (Print)1064-3745

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    Walsh, E. P., Lamont, D., Beattie, K. A., & Stark, M. J. R. (2007). Proteins interacting with Saccharomyces cerevisiae type 1 protein phosphatase catalytic subunit identified by single-step affinity purification and mass spectrometry. In G. Moorhead (Ed.), Protein phosphatase protocols (pp. 235-246). (Methods in Molecular Biology; Vol. 365). Humana Press. https://doi.org/10.1385/1-59745-267-X:235