Abstract
This method describes an in vitro assay for measuring INO1 enzyme activity (the conversion of glucose 6-phosphate to myo-inositol 3-phosphate) in cell-free extracts. The method was first described for Plasmodium falciparum cells in MacRae et al. (2014) and consists of two parts: Part 1 describes the assay itself while part 2 describes analysis of the myo-inositol 3-phosphate product using gas chromatography-mass spectrometry (GC-MS).
Materials and Reagents
Equipment
Software
Procedure
Note: An overview of the whole procedure can be seen in the accompanying figure. Figure 1. Inositol 3-phosphate synthase assay: overview
Recipes
Acknowledgments
This work was supported by a project grants from the National Health and Medical Research Council of Australia (NHMRC). M.J.M is an NHMRC Principal Research Fellow and J.I.M. was supported by a Royal Society Travelling Fellowship.
References
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