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A simple and highly sensitive radioenzymatic assay for lysophosphatidic acid quantification
J S Saulnier-Blache1, A Girard, M F Simon
1INSERM U317, Institut Louis Bugnard, Université Paul Sabatier, CHU Rangueil, 31403 Toulouse, Cedex 04, France.
Abstract:
The objective of the present work was to develop a simple and sensitive radioenzymatic assay to quantify lysophosphatidic acid (LPA). For that, a recombinant rat LPA acid acyltransferase (LPAAT) produced in Escherichia coli was used. In the presence of [(14)C]oleoyl-CoA, LPAAT selectively catalyzes the transformation of LPA and alkyl-LPA into [(14)C]phosphatidic acid. Acylation of LPA was complete and linear from 0 to 200 pmol with a minimal detection of 0.2 pmol. This method was used to quantify LPA in butanol-extracted lipids from bovine sera, as well as from human and mouse plasma. This radioenzymatic assay represents a new, simple, and highly sensitive method to quantify LPA in various biological fluids.