Abstract
Phosphatidylinositol (PtdIns) synthase is a lipid-synthesizing enzyme responsible for the synthesis of the phospholipid, PtdIns. Its enzymatic properties have been studied in in vitro assays using either membrane-enriched fractions or the purified protein in reconstituted lipid vesicles as a source of enzyme. More recently the specificities in terms of substrate preferences have also been studied using the recombinant protein expressed in Escherichia coli. This chapter deals with the purification of membranes as a source of PtdIns synthase before focusing on the in vitro assays of the enzymatic activities of the protein and, briefly, on the analysis of the product.
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Acknowledgments
The authors wish to thank E. Ruelland for his careful reading of the manuscript. Financial support from the Université Pierre et Marie Curie via the French Ministère de l’Enseignement Supérieur et de la Recherche, as well as from the C.N.R.S., is gratefully acknowledged.
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Collin, S., Cochet, F. (2013). Phosphatidylinositol Synthase Activity from Plant Membrane Fractions and from E. coli- Expressed Recombinant Peptides. In: Munnik, T., Heilmann, I. (eds) Plant Lipid Signaling Protocols. Methods in Molecular Biology, vol 1009. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-62703-401-2_14
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DOI: https://doi.org/10.1007/978-1-62703-401-2_14
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