A direct nonchromatographic assay for 1-acyl-sn-glycerol-3-phosphate acyltransferase

Rajasekharan, Ram ; Ray, Tapas K. ; Cronan Jr., John E. (1988) A direct nonchromatographic assay for 1-acyl-sn-glycerol-3-phosphate acyltransferase Analytical Biochemistry, 173 (2). pp. 376-382. ISSN 0003-2697

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Official URL: http://www.sciencedirect.com/science/article/pii/0...

Related URL: http://dx.doi.org/10.1016/0003-2697(88)90202-3


1-Acyl-sn-glycerol-3-phosphate acyltransferase (also called lysophosphatidic acid acyltransferase) which catalyzes the acylation of 1-acyl-sn-glycerol-3-phosphate to phosphatidic acid is generally assayed by the use of a radioactive substrate followed by a time-consuming chromatographic separation of substrate and product. We report a direct and highly sensitive nonchromatographic assay for this enzyme based on the ability of Escherichia coli alkaline phosphatase to dephosphorylate 1-acyl-sn-glycerol-3-phosphate but not phosphatidic acid. This selective hydrolysis coupled with the use of 32P-labeled 1-acyl-sn-glycerol-3-phosphate as substrate permits measurement of the product, 32P-labeled phosphatidic acid by solvent extraction or precipitation. We also report a series of enzymatic reactions for the efficient conversion of 32Pi to 32P-labeled 1-acyl-sn-glycerol-3-phosphate.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:Acyltransferase; Lysophosphatidic Acid; Acylation; Assay; Phosphatidic Acid
ID Code:55066
Deposited On:17 Aug 2011 12:16
Last Modified:17 Aug 2011 12:16

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