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

doi:10.3791/57646

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

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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 language	English (US)
Journal	Journal of visualized experiments : JoVE
Issue number	141
DOIs	https://doi.org/10.3791/57646
State	Published - 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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Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay. / Alizadeh, Javad; Shojaei, Shahla; da Silva Rosa, Simone; Rezaei Moghadam, Adel; Zeki, Amir A.; Hashemi, Mohammad; Los, Marek J.; Gordon, Joseph W.; Ghavami, Saeid.

In: Journal of visualized experiments : JoVE, No. 141, 11.11.2018.

Research output: Contribution to journal › Article › peer-review

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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.",

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AU - da Silva Rosa, Simone

AU - Rezaei Moghadam, Adel

AU - Zeki, Amir A.

AU - Hashemi, Mohammad

AU - Los, Marek J.

AU - Gordon, Joseph W.

AU - Ghavami, Saeid

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AB - 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.

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

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