A simple and highly sensitive radioenzymatic assay for lysophosphatidic acid quantification.
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.
Saulnier-Blache
J S
JS
INSERM U317, Institut Louis Bugnard, Université Paul Sabatier, CHU Rangueil, 31403 Toulouse, Cedex 04, France.
Girard
A
A
Simon
M F
MF
Lafontan
M
M
Valet
P
P
D000217:Acyltransferases / Q000737:chemistry; D000818:Animals; D002855:Chromatography, Thin Layer; D008246:Lysophospholipids / Q000032:analysis*; D008722:Methods; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D012680:Sensitivity and Specificity
D008246:Lysophospholipids; D011994:Recombinant Proteins; D000217:Acyltransferases; C018764:2-acylglycerophosphate acyltransferase