Tumaney, Ajay W and Shekar, Sunil and Rajasekharan, Ram (2001) Identification, Purification, and Characterization of Monoacylglycerol Acyltransferase from Developing Peanut Cotyledons. In: Journal of Biological Chemistry, 276 (14). pp. 10847-10852.
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Abstract
Biosynthesis of diacylglycerols in plants occurs mainly through the acylation of lysophosphatidic acid in the microsomal membranes. Here we describe the first identification of diacylglycerol biosynthetic activity in the soluble fraction of developing oilseeds. This activity was NaF-insensitive and acyl-CoA-dependent. Diacylglycerol formation was catalyzed by monoacylglycerol (MAG) acyltransferase (EC 2.3.1.22) that transferred an acyl moiety from acyl-CoA to MAG. The enzyme was purified by successive chromatographic separations on octyl-Sepharose, blue-Sepharose, Superdex-75, and palmitoyl-CoA-agarose to apparent homogeneity from developing peanut (Arachis hypogaea) cotyledons. The enzyme was purified to 6,608-fold with the final specific activity of 15.86 nmol $min^{-1} mg^{-1}$. The purified enzyme was electrophoretically homogeneous, and its molecular mass was 43,000 daltons. The purified MAG acyltransferase was specific for MAG and did not utilize any other acyl acceptor such as glycerol, glycerol-3-phosphate, lysophosphatidic acid, and lysophosphatidylcholine. The $K_m$ values for 1-palmitoylglycerol and 1-oleoylglycerol were 16.39 and 5.65 $\mu$M, respectively. The $K_m$ values for 2-monoacylglycerols were 2- to 4-fold higher than that of the corresponding 1-monoacylglycerol. The apparent $K_m$ values for palmitoyl-, stearoyl-, and oleoyl-CoAs were 17.54, 25.66, and 9.35 $\mu$M, respectively. Fatty acids, phospholipids, and sphingosine at low concentrations stimulated the enzyme activity. The identification of MAG acyltransferase in oilseeds suggests the presence of a regulatory link between signal transduction and synthesis of complex lipids in plants.
Item Type: | Journal Article |
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Publication: | Journal of Biological Chemistry |
Publisher: | American Society for Biochemistry and Molecular Biology |
Additional Information: | Copyright of this article belongs to American Society for Biochemistry and Molecular Biology |
Keywords: | Enzymology;Biochemistry;Molecular Biophysics;Botany;Methods and Techniques |
Department/Centre: | Division of Biological Sciences > Biochemistry |
Date Deposited: | 26 Oct 2007 |
Last Modified: | 19 Sep 2010 04:40 |
URI: | http://eprints.iisc.ac.in/id/eprint/12174 |
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