Methods in the study of PTEN structure: X-ray crystallography and hydrogen deuterium exchange mass spectrometry

Glenn R. Masson, John E. Burke, Roger L. Williams (Lead / Corresponding author)

    Research output: Chapter in Book/Report/Conference proceedingChapter

    3 Citations (Scopus)

    Abstract

    Despite its small size and deceptively simple domain organization, PTEN remains a challenging structural target due to its N- and C-terminal intrinsically disordered segments, and the conformational heterogeneitycaused by phosphorylation of its C terminus. Using hydrogen/deuterium exchange mass spectrometry (HDX-MS), it is possible to probe the conformational dynamics of the disordered termini, and also to determine how PTEN binds to lipid membranes. Here, we describe how to purify recombinant, homogenously dephosphorylated PTEN from a eukaryotic system for subsequent investigation with HDX-MS or crystallography.

    Original languageEnglish
    Title of host publicationPTEN
    Subtitle of host publicationMethods in Molecular Biology
    EditorsLeonardo Salmena, Vuk Stambolic
    PublisherHumana Press
    Pages215-230
    Number of pages16
    ISBN (Electronic)978-1-4939-3299-3
    ISBN (Print)978-1-4939-3297-9
    DOIs
    Publication statusPublished - 2016

    Publication series

    NameMethods in Molecular Biology
    Volume1388
    ISSN (Print)1064-3745

    Keywords

    • HDX-MS
    • Insect cell expression
    • Liposomeb preparation
    • Mass spectrometry
    • Protein purification

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics

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