Purification and Characterization of 1-O-Acylceramide Synthase, a Novel Phospholipase A2 with Transacylase Activity

Abe, Akira; Shayman, James A.

doi:10.1074/jbc.273.14.8467

Cited by 88 publications

(87 citation statements)

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“…Time courses for the release of oleic acid and the formation of 2-oleoyl-lysoPC are shown in C and D, respectively. from other phospholipases that lack transacylase activity (1)(2)(3)(4)8). The transacylase activity may also be useful to resolve the positional specificity of Lpla2.…”

Section: /2mentioning

confidence: 99%

“…Transacylase activity of the soluble fraction obtained from Lpla2-overexpressing MDCK cells Lpla2 has dual enzyme activities: PLA 2 and transacylase activities (1,2). The most common assay for transacylase activity by Lpla2 measures the transacylation of a fatty acid between phospholipid (PC and PE) and a short-chain ceramide such as NAS as an acceptor.…”

Section: /2mentioning

confidence: 99%

“…This phospholipase is calcium-independent, is localized to lysosomes, has an acidic pH optimum, and transacylates short-chain ceramides (1-3). Lpla2 was purified from bovine brain and characterized as a water-soluble glycoprotein consisting of a single peptide chain of 45 kDa (2). The primary structure of Lpla2, deduced from DNA sequences coding Lpla2, is highly conserved between mammals, including mouse, rat, human, and cow (3,4).…”

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Positional specificity of lysosomal phospholipase A2

Abe

Hiraoka

Shayman

2006

Journal of Lipid Research

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Lysosomal phospholipase A 2 (Lpla2) is highly expressed in alveolar macrophages and may mediate the phospholipid metabolism of surfactant. Studies on the properties of this phospholipase are consistent with the presence of both phospholipase A 1 and phospholipase A 2 activities. These activities were studied through the production of O-acyl compounds, produced by the transacylase activity of Lpla2. Liposomes containing POPC and N-acetylsphingosine (NAS) were incubated with the soluble fraction obtained from MDCK cells stably transfected with the mouse Lpla2 gene. Two 1-O-acyl-NASs, 1-O-palmitoyl-NAS and 1-O-oleoyl-NAS, were produced by Lpla2. The formation rate of 1-O-oleoyl-NAS was 2.5-fold that of 1-O-palmitoyl-NAS. When 1-oleoyl-2-palmitoyl-sn-glycero-3-phosphocholine (OPPC) was used, the formation rate of 1-O-oleoyl-NAS was 5-fold higher than that of 1-O-palmitoyl-NAS. Thus, Lpla2 can act on acyl groups at both sn-1 and sn-2 positions of POPC and OPPC. When 1-palmitoyl-2-unsaturated acyl-sn-glycero-3-phosphocholines were used as acyl donors, the transacylation of the acyl group from the sn-2 position to NAS was preferred to that of the palmitoyl group from the sn-1 position. An exception was observed for 1-palmitoyl-2-arachidonoyl-snglycero-3-phosphocholine (PAPC), for which the formation rate of 1-O-palmitoyl-NAS from PAPC was 4-fold greater than that of 1-O-arachidonoyl-NAS. Thus, Lpla2 has broad positional specificity for the sn-1 and sn-2 acyl groups in phosphatidylcholine and phosphatidylethanolamine.-Abe, A., M. Hiraoka, and J. A. Shayman. Positional specificity of lysosomal phospholipase A 2 .

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Section: /2mentioning

confidence: 99%

Section: /2mentioning

confidence: 99%

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Positional specificity of lysosomal phospholipase A2

Abe

Hiraoka

Shayman

2006

Journal of Lipid Research

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“…The specific activity of lysophospholipase was significantly lower than that of ACS. Purified ACS from bovine brain has weak enzyme activity as a phospholipase A 1 (4). Therefore, the lysophospholipase activity observed in recombinant LLPL probably reflects the minor phospholipase A 1 activity of ACS.…”

Section: Fig 4 Llpl Has Both Transacylase and Phospholipase Activitmentioning

confidence: 99%

“…The new enzyme, named 1-O-acylceramide synthase (ACS), was purified from bovine brain and further characterized (4). ACS is a water-soluble glycoprotein with a molecular mass of 45 kDa and a single polypeptide chain, which specifically binds to concanavalin A-conjugated agarose.…”

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Cloning and Characterization of a Lysosomal Phospholipase A2, 1-O-Acylceramide Synthase

Hiraoka¹,

Abe²,

Shayman

2002

Journal of Biological Chemistry

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107

113

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Recently, a novel enzyme, 1-O-acylceramide synthase (ACS), was purified and characterized from bovine brain. This enzyme has both calcium-independent phospholipase A 2 and transacylase activities. The discovery of this enzyme led us to propose a new pathway for ceramide metabolism in which the sn-2-acyl group of either phosphatidylethanolamine or phosphatidylcholine is transferred to the 1-hydroxyl group of ceramide. In this study, the partial amino acid sequences from the purified enzyme revealed that the enzyme contains amino acid sequences identical to those of human lecithin:cholesterol acyltransferase-like lysophospholipase (

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