An evaluation of strategies available for the identification of GTP-binding proteins required in intracellular signalling pathways

Greg J. Barritt, Roland B. Gregory

    Research output: Contribution to journalReview articlepeer-review

    22 Citations (Scopus)

    Abstract

    Strategies which can be used to elucidate the nature of a GTP-binding regulatory protein (G-protein) involved in an intracellular pathway of interest in the complex environment of the cell are described and evaluated. A desirable strategy is considered to be one in which the first stage indicates a requirement for one or more G-proteins, provides information on whether a monomeric, trimeric or other type of G-protein is involved, and gives some idea of the G protein sub-class. In the second stage the specific G-protein involved is identified. Approaches available for investigations in the first stage include the use of analogues of GTP and GDP, AlF4̄, inhibitors of G-protein isoprenylation, bacterial toxins which covalently modify G-proteins, and the introduction of a purified GDP dissociation inhibitor, GDP exchange and/or GTP-ase activating protein. Identification of the specific G-protein in the second stage can be achieved using anti G-protein antibodies, G-protein- or receptor-derived peptides, antisense G protein RNA and over-expressed, constitutively-active or dominant-negative G-protein mutants. The correct interpretation of results obtained with GTP and GDP analogues and AlF4̄ in the first stage is complex and often difficult, and requires a thorough understanding of the functions and mechanisms of activation of G-proteins. Nevertheless, it is important to reach the correct conclusion at this stage since considerable time and expense are usually required for investigations in the second stage.

    Original languageEnglish
    Pages (from-to)207-218
    Number of pages12
    JournalCellular Signalling
    Volume9
    Issue number3-4
    DOIs
    Publication statusPublished - May 1997

    Keywords

    • AlF̄
    • Anti G-protein antibodies
    • Antisense RNA
    • G-protein mutants
    • G-protein peptides
    • GTPγS
    • GTP-binding regulatory proteins
    • Strategy for identification

    Fingerprint Dive into the research topics of 'An evaluation of strategies available for the identification of GTP-binding proteins required in intracellular signalling pathways'. Together they form a unique fingerprint.

    Cite this