Abstract
The methods described in this chapter are designed to measure the hydrolysis of guanosine triphosphate (GTP) to inorganic phosphate (Pi) and guanosine diphosphate (GDP), a reaction catalyzed by the GTPase enzymes (EC 3.6.1.-). The theory behind the experimental design involves using [γ-32P]GTP as a marker, whereby any GTPase-induced hydrolysis will result in the 32P label appearing as 32P, and as unhydrolyzed [γ-32P]GTP. It was originally described by Cassel and Selinger (1). 32P, must be separated from [γ-32P]GTP, and then can be easily quantitated by using a liquid scintillation counter. The amount of 32Pi is directly proportional to the amount of [γ-32P]GTP hydrolyzed, and, therefore, proportional to the activity of GTPases in the preparation.
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Sweeney, M. (1995). Measurement of the GTPase Activity of Signal-Transducing G-Proteins in Neuronal Membranes. In: Kendall, D.A., Hill, S.J. (eds) Signal Transduction Protocols. Methods in Molecular Biology™, vol 41. Humana Press. https://doi.org/10.1385/0-89603-298-1:51
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DOI: https://doi.org/10.1385/0-89603-298-1:51
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