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

DOI :

10.3791/57646-v

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November 11th, 2018

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9,592 Views

Javad Alizadeh^1,2, Shahla Shojaei^1,2,3, Simone da Silva Rosa¹, Adel Rezaei Moghadam^1,2, Amir A. Zeki⁴, Mohammad Hashemi⁵, Marek J. Los^6,7,8, Joseph W. Gordon^1,9, Saeid Ghavami^1,2,10

¹Department of Human Anatomy and Cell Science, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of Manitoba, ²Biology of Breathing Theme, Children Hospital Research Institute of Manitoba, University of Manitoba, ³Department of Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, ⁴Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, Center for Comparative Respiratory Biology and Medicine, ⁵Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, ⁶Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, ⁷Centre de biophysique moléculaire - UPR 4301, Centre national de la recherche scientifique (CNRS) CS80054, ⁸LinkoCare Life Sciences AB, ⁹College of Nursing and Children's Hospital Research Institute of Manitoba, Rady Faculty of Health Sciences, University of Manitoba, ¹⁰Health Policy Research Center, Institute of Health, Shiraz University of Medical Sciences

Chapters

Here we describe a protocol to investigate the prenylation and guanosine-5'-triphosphate (GTP)-loading of Rho GTPase. This protocol consists of two detailed methods, namely membrane fractionation and a GTPase-linked immunosorbent assay. The protocol can be used for measuring the prenylation and GTP loading of different other small GTPases.

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