Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay

Javad Alizadeh, Shahla Shojaei, Simone da Silva Rosa, Adel Rezaei Moghadam, Amir A. Zeki, Mohammad Hashemi, Marek J. Los, Joseph W. Gordon, Saeid Ghavami

    Research output: Contribution to journalArticle

    2 Scopus citations

    Abstract

    The Rho GTPase family belongs to the Ras superfamily and includes approximately 20 members in humans. Rho GTPases are important in the regulation of diverse cellular functions, including cytoskeletal dynamics, cell motility, cell polarity, axonal guidance, vesicular trafficking, and cell cycle control. Changes in Rho GTPase signaling play an essential regulatory role in many pathological conditions, such as cancer, central nervous system diseases, and immune system-dependent diseases. The posttranslational modification of Rho GTPases (i.e., prenylation by mevalonate pathway intermediates) and GTP binding are key factors which affect the activation of this protein. In this paper, two essential and simple methods are provided to detect a broad range of Rho GTPase prenylation and GTP binding activities. Details of the technical procedures that have been used are explained step by step in this manuscript.

    Original languageEnglish (US)
    JournalJournal of visualized experiments : JoVE
    Issue number141
    DOIs
    StatePublished - Nov 11 2018

    ASJC Scopus subject areas

    • Neuroscience(all)
    • Chemical Engineering(all)
    • Biochemistry, Genetics and Molecular Biology(all)
    • Immunology and Microbiology(all)

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  • Cite this

    Alizadeh, J., Shojaei, S., da Silva Rosa, S., Rezaei Moghadam, A., Zeki, A. A., Hashemi, M., Los, M. J., Gordon, J. W., & Ghavami, S. (2018). Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay. Journal of visualized experiments : JoVE, (141). https://doi.org/10.3791/57646