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Detection of Small Gtpase Prenylation and Gtp Binding Using Membrane Fractionation and Gtpase-Linked Immunosorbent Assay Publisher Pubmed



Alizadeh J1, 2 ; Shojaei S1, 2, 3 ; Rosa SDS1 ; Moghadam AR1, 2 ; Zeki AA4 ; Hashemi M5 ; Los MJ6, 7, 8 ; Gordon JW1, 9 ; Ghavami S1, 2, 10
Authors
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Authors Affiliations
  1. 1. Department of Human Anatomy and Cell Science, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of Manitoba, Canada
  2. 2. Biology of Breathing Theme, Children Hospital Research Institute of Manitoba, University of Manitoba, Canada
  3. 3. Department of Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Canada
  4. 4. Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, Center for Comparative Respiratory Biology and Medicine, Canada
  5. 5. Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Canada
  6. 6. Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Canada
  7. 7. Centre de biophysique moleculaire-UPR 4301, Centre national de la recherche scientifique (CNRS), CS80054, Canada
  8. 8. LinkoCare Life Sciences AB, Canada
  9. 9. College of Nursing and Children's Hospital Research Institute of Manitoba, Rady Faculty of Health Sciences, University of Manitoba, Canada
  10. 10. Health Policy Research Center, Institute of Health, Shiraz University of Medical Sciences, Canada

Source: Journal of Visualized Experiments Published:2018


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. © 2018 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License.
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