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cDNA Cloning and Expression of Murine 1-Acyl-sn-glycerol-3-phosphate Acyltransferase

https://doi.org/10.1006/bbrc.1997.7214Get rights and content

Abstract

1-Acyl-sn-glycerol-3-phosphate (1-AGP), also known as lysophosphatidic acid (LPA), is an intermediate ofde novobiosynthesis of glycerophospholipids and triacylglycerol. LPA is also attracting much attention because of its growth stimulating effects. Here we report cloning of murine cDNA encoding 1-AGP acyltransferase (1-AGPAT), which converts LPA into phosphatidic acid by incorporating acyl moiety atsn-2 position. The cDNA contains an open reading frame coding for 285 amino acids, with highly hydrophobic regions in the N-terminal half. The Northern blot analysis using various mouse tissues revealed ubiquitous expressions of two transcripts. The cDNA was expressed inEscherichia coliJC201, aplsCmutant strain deficient in the 1-AGPAT activity, and the enzyme properties were examined. The enzyme utilized both saturated and unsaturated acyl-CoA as an acyl-donor, while it utilized LPA but not other lysophospholipids as an acceptor.

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    Abbreviations used: 1-AGP, 1-acyl-sn-glycerol-3-phosphateLPA, lysophosphatidic acid; 1-AGPAT, 1-AGP acyltransferase; GP,sn -glycerol-3-phosphate; GPAT, GP acyltransferase; E. coli, Escherichia coli;; EST, expressed sequence tag; LPC, lysophosphatidylcholine; lysoPAF, lyso-platelet-activating factor; ORF, open reading frame; IPTG, isopropyl-β-D-thiogalactopyranoside;

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