Subcellular distribution and properties of acyl/alkyl dihydroxyacetone phosphate reductase in rodent livers

Ghosh, Mridul K.; Hajra, Amiya K.

doi:10.1016/0003-9861(86)90245-6

Cited by 40 publications

(20 citation statements)

References 36 publications

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“…These results agree well with the labeling pattern observed when using 32 P i (Table III). The addition of AG to the medium restored the labeling of plasmenylethanolamine to near wild-type values, suggesting that the steps downstream of acyl/alkyl-DHAP re- [9, …”

Section: Identification Of the Enzymatic Lesion In Faak1b As A Defimentioning

confidence: 94%

“…Accumulation and Distribution of [9, H]Hexadecanol-Cells were plated into sterile, 20-ml glass scintillation vials, at 2 ϫ 10 5 cells/vial, in 1 ml of F12c and allowed to attach overnight at 37°C. The following day, 0.5 ml of F12c containing 15 M [9,10-3 H]hexadecanol (2 ϫ 10 6 cpm/vial) was added to each vial (final hexadecanol concentration of 5 M), and the cells were incubated at 37°C for 1.5 or 3 h. Medium was removed, the cells were washed 2 times with 3 ml of F12c followed by one wash with 3 ml of PBS.…”

Section: Methodsmentioning

confidence: 99%

“…Alkyl-DHAP synthase activity was assayed by measuring the incorporation of [1][2][3][4][5][6][7][8][9][10][11][12][13][14] C]hexadecanol into alkyl-DHAP in the presence of 1-acyl-DHAP, as described by Davis and Hajra (14). Acyl/alkyl-DHAP reductase was monitored by measuring the incorporation of label from B- [4-3 H]NADPH into chloroform-soluble counts in the presence of 1-Ohexadecyl-DHAP (30) or 1-palmitoyl-DHAP (2).…”

Section: Methodsmentioning

confidence: 99%

“…Acyl/alkyl-DHAP reductase is a membrane-bound enzyme that has been localized to the cytosolic face of both the peroxisomal (8) and microsomal membranes (9). Interestingly, in both human (10,11) and rodent (12,13) cells that do not assemble peroxisome properly, other activities associated with the peroxisomal membrane, such as DHAP acyltransferase and alkyl-DHAP synthase, are severely decreased, but acyl/alkyl-DHAP reductase activity remains unaffected.…”

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confidence: 99%

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An Animal Cell Mutant with a Deficiency in Acyl/Alkyl-dihydroxyacetone-phosphate Reductase Activity

James¹,

Lake²,

Hajra³

et al. 1997

Journal of Biological Chemistry

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In the accompanying paper (James, P. F., and Zoeller, R. A. (1997) J. Biol. Chem. 272, 23532-23539), we reported the isolation of a series of mutants from the fibroblastlike cell line, CHO-K1, that are deficient in the incorporation of the long chain fatty alcohol, hexadecanol, into complex lipids. All but one of these mutants, FAA.K1B, were deficient in long-chain-fatty alcohol oxidase (FAO) activity. We have further characterized this FAO ؉ isolate. FAA.K1B cells displayed a 40% decrease in [9,10-3 H]hexadecanol uptake when compared with the parent strain. Although incorporation of hexadecanol into the phospholipid fraction was decreased by 52%, the cells accumulated label in alkylglycerol (20-fold over wild type). The increase in 1-alkylglycerol labeling corresponded to a 4-fold increase in alkylglycerol mass. Short term labeling with 32 P i showed a 45-50% decrease in overall phospholipid biosynthesis in FAA.K1B. Both diacyl-and ether-linked species were affected, suggesting a general defect in phospholipid biosynthesis. Mutant cells were able to partially compensate for the decreased biosynthesis by decreasing the turnover of the phospholipid pools. The primary lesion in FAA.K1B was identified as a 95% reduction in acyl/alkyl-dihydroxyacetone-phosphate reductase activity. Whole cell homogenates from FAA.K1B were unable to reduce either acyldihydroxyacetone phosphate (DHAP) or alkyl-DHAP, supporting the notion that the reduction of these two compounds is catalyzed by a single enzyme. These data suggest that the biosynthesis of diacyl phospholipids, in Chinese hamster ovary cells, begins with the acylation of dihydroxyacetone phosphate as well as glycero-3-phosphate and that the "DHAP pathway" contributes significantly to diacyl glycerolipid biosynthesis. Also, the severe reduction in acyl/alkyl-DHAP reductase activity in FAA.K1B resulted in only a moderate decrease in ether lipid biosynthesis. These latter data together with the observed increase in alkylglycerol levels support the existence of a shunt pathway that is able to partially bypass the enzymatic lesion.The formation of phosphatidic acid is required for the formation of diacyl glycerolipids in both bacterial and animal cells. In bacterial systems, this is initiated with the reduction of dihydroxyacetone phosphate (DHAP), 1 a product of glycolysis. The resulting sn-glycero-3-phosphate is acylated sequentially at the sn-1 and sn-2 positions to form phosphatidic acid. In animal cells, dihydroxyacetone phosphate (DHAP) can be acylated directly. In this "DHAP pathway," the ketone at the sn-2 position of the resulting sn-1-acyl-DHAP must be reduced, by acyl/alkyl-DHAP reductase (1), prior to further acylation. Purified acyl/ alkyl-DHAP reductase also catalyzes the reduction of the ether-linked sn-1-alkyl-DHAP (2), a reaction that is required in the synthesis of ether-linked glycerolipids, such as plasmenylethanolamine and glyceryl ether diesters (1). While the acylation of DHAP is accepted as the first step in ether-linked glycerolipids, the importance o...

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Section: Identification Of the Enzymatic Lesion In Faak1b As A Defimentioning

confidence: 94%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

An Animal Cell Mutant with a Deficiency in Acyl/Alkyl-dihydroxyacetone-phosphate Reductase Activity

James¹,

Lake²,

Hajra³

et al. 1997

Journal of Biological Chemistry

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“…Regulation of these enzymes by oxygen might be related to their subcellular localization. Zinser and coworkers (42) (8).…”

Section: Resultsmentioning

confidence: 99%

The acyl dihydroxyacetone phosphate pathway enzymes for glycerolipid biosynthesis are present in the yeast Saccharomyces cerevisiae

et al. 1992

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The presence of the acyl dihydroxyacetone phosphate (acyl DHIAP) pathway in yeasts was investigated by examining three key enzyme activities of this pathway in Saccharomyces cerevisiae. In the total membrane fraction of S. cerevisiae, we confirmed the presence of both DHAP acyltransferase (DHAPAT; Km = 1.27 mM; Vmax = 5.9 nmol/min/mg of protein) and sn-glycerol 3-phosphate acyltransferase (GPAT; Km = 0.28 mM; Vmax = 12.6 nmol/min/mg of protein). The properties of these two acyltransferases are similar with respect to thermal stability and optimum temperature of activity but differ with respect to pH optimum (6.5 for GPAT and 7.4 for DHAPAT) and sensitivity toward the sulfhydryl blocking agent N-ethylmaleimide. Total membrane fraction of S. cerevisiae also exhibited acyl/alkyl DHAP reductase (EC 1.1.1.101) activity, which has not been reported previously. The reductase has a Vm, of 3.8 nmol/min/mg of protein for the reduction of hexadecyl DHLAP (Km = 15 ,uM) by NADPH (Km = 20 ,uM). Both acyl DHAP and alkyl DHAP acted as substrates. NADPH was the specific cofactor. Divalent cations and N-ethylmaleimide inhibited the enzymatic reaction.Reductase activity in the total membrane fraction from aerobically grown yeast cells was twice that from anaerobically grown cells. Similarly, DHAPAT and GPAT activities were also greater in aerobically grown yeast cells. The presence of these enzymes, together with the absence of both ether glycerolipids and the ether lipid-synthesizing enzyme (alkyl DHIAP synthase) in S. cerevisiae, indicates that non-ether glycerolipids are synthesized in this organism via the acyl DHAP pathway.Biosynthesis of phosphatidic acid, the precursor of glycerolipids in all organisms, proceeds via two different routes. In the glycerol phosphate pathway, phosphatidic acid is biosynthesized via the stepwise enzymatic acylation of snglycerol 3-phosphate (GP) by long-chain fatty acyl coenzyme A's (acyl-CoAs). In animals, phosphatidate is also biosynthesized by an alternate pathway, the acyl dihydroxyacetone phosphate (acyl DHAP) pathway. In this pathway, DHAP is acylated by a long-chain fatty acyl-CoA to form acyl DHAP. This compound is then reduced to 1-acyl GP, which is further acylated to yield phosphatidate (Fig. 1). The GP pathway is universal to all organisms, including yeasts (19,27). In contrast, the acyl DHAP pathway is absent in bacteria and plants (11, 41). In animals, the acyl DHAP pathway is obligatory for ether lipid biosynthesis; the ether bond is synthesized by the substitution of a long-chain alcohol for the acyl group of acyl DHAP, with the release of free fatty acid to form alkyl DHAP (13). The enzymatic conversion of alkyl DHAP to glycerol ether lipids proceeds via reduction of alkyl DHAP to 1-alkyl GP followed by acylation (13). The importance of the acyl DHAP pathway for the biosynthesis of non-ether glycerolipids is controversial (6,29,30,33,34,35). Except for some anaerobic bacteria which contain plasmalogens but not alkyl glycerol ethers, both aerobic bacteria and facultative anaerobe...

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From peroxisomal disorders to common neurodegenerative diseases – the role of ether phospholipids in the nervous system

2017

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The emerging diverse roles of ether (phospho)lipids in nervous system development and function in health and disease are currently attracting growing interest. Plasmalogens, a subgroup of ether lipids, are important membrane components involved in vesicle fusion and membrane raft composition. They store polyunsaturated fatty acids and may serve as antioxidants. Ether lipid metabolites act as precursors for the formation of glycosyl-phosphatidyl-inositol anchors; others, like platelet-activating factor, are implicated in signaling functions. Consolidating the available information, we attempt to provide molecular explanations for the dramatic neurological phenotype in ether lipid-deficient human patients and mice by linking individual functional properties of ether lipids with pathological features. Furthermore, recent publications have identified altered ether lipid levels in the context of many acquired neurological disorders including Alzheimer's disease (AD) and autism. Finally, current efforts to restore ether lipids in peroxisomal disorders as well as AD are critically reviewed.

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Subcellular distribution and properties of acyl/alkyl dihydroxyacetone phosphate reductase in rodent livers

Cited by 40 publications

References 36 publications

An Animal Cell Mutant with a Deficiency in Acyl/Alkyl-dihydroxyacetone-phosphate Reductase Activity

An Animal Cell Mutant with a Deficiency in Acyl/Alkyl-dihydroxyacetone-phosphate Reductase Activity

The acyl dihydroxyacetone phosphate pathway enzymes for glycerolipid biosynthesis are present in the yeast Saccharomyces cerevisiae

From peroxisomal disorders to common neurodegenerative diseases – the role of ether phospholipids in the nervous system

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