Formation of diacylglycerol by a phospholipase D-phosphatidate phosphatase pathway specific for phosphatidylcholine in endothelial cells
The conversion of phosphatidylcholine (PC) to diacylglycerol (DAG) was studied in sonicated endothelial cells and in subcellular fractions in the presence of 0.05% Triton X-100 and 2 mM EDTA. DAG formation occurred predominantly in an organelle fraction that sedimented at 15 000 x g. In parallel rea...
Ausführliche Beschreibung
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| Autor*in: |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
1988 |
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| Reproduktion: |
Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 |
|---|---|
| Übergeordnetes Werk: |
in: Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism - Amsterdam : Elsevier, 962(1988), 3, Seite 282-296 |
| Übergeordnetes Werk: |
volume:962 ; year:1988 ; number:3 ; pages:282-296 |
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| 520 | |a The conversion of phosphatidylcholine (PC) to diacylglycerol (DAG) was studied in sonicated endothelial cells and in subcellular fractions in the presence of 0.05% Triton X-100 and 2 mM EDTA. DAG formation occurred predominantly in an organelle fraction that sedimented at 15 000 x g. In parallel reactions with exogenous 1-oleoyl-2-[^3H]oleoyl-PC (sn-2-[^3H]DOPC) and phosphatidyl[^3H]choline ([choline-^3H]PC), [^3H]DAG was formed by a reaction pathway in which [^3H]choline was the only product derived from [choline-^3H]PC. [^3H]Choline was not formed secondarily from [^3H]glycerophosphocholine or [^3H]phosphocholine. Small amounts of [^3H]phosphatidate ([^3H]PA) were isolated from reactions with sn-2-[^3H]DOPC at short incubation times, and substantial PA phosphatase activity was demonstrated. These data, taken together, supported a phospholipase D-PA phosphatase pathway of DAG formation. Kinetic data established that the low ratio of [^3H]PA/[^3H]DAG formed in reactions with sn-2-[^3H]DOPC was due to a 15-fold higher V"m"a"x and 7-fold lower apparent K"m of the PA phosphatase. The [^3H]PA/[^3H]DAG product ratio was increased by addition of unlabeled PA or by selective extraction of phospholipase D with Triton X-100. The characteristics of the phospholipase D indicated a unique enzyme. Activity was optimal in the presence of EDTA and was almost totally dependent upon Triton X-100. The pH profile displayed a peak at 7.0. Of particular significance was the stringent substrate specificity. Phosphatidylinositol was not hydrolyzed, and activities towards phosphatidylethanolamine and sphingomyelin were at most 30- to 50-fold lower than those towards PC. Phospholipase D and PA phosphatase were identified in a number of rat tissues and other cells. The highest activities of phospholipase D were present in lung and endothelial cells. Phospholipase D was partially purified from rat lung by Triton X-100 extraction and anion exchange chromatography. When linked with PA phosphatase, the phospholipase D could initiate a pathway of DAG formation that is highly specific for PC. | ||
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(DE-627)NLEJ185922686 (DE-599)GBVNLZ185922686 DE-627 ger DE-627 rakwb eng Formation of diacylglycerol by a phospholipase D-phosphatidate phosphatase pathway specific for phosphatidylcholine in endothelial cells 1988 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The conversion of phosphatidylcholine (PC) to diacylglycerol (DAG) was studied in sonicated endothelial cells and in subcellular fractions in the presence of 0.05% Triton X-100 and 2 mM EDTA. DAG formation occurred predominantly in an organelle fraction that sedimented at 15 000 x g. In parallel reactions with exogenous 1-oleoyl-2-[^3H]oleoyl-PC (sn-2-[^3H]DOPC) and phosphatidyl[^3H]choline ([choline-^3H]PC), [^3H]DAG was formed by a reaction pathway in which [^3H]choline was the only product derived from [choline-^3H]PC. [^3H]Choline was not formed secondarily from [^3H]glycerophosphocholine or [^3H]phosphocholine. Small amounts of [^3H]phosphatidate ([^3H]PA) were isolated from reactions with sn-2-[^3H]DOPC at short incubation times, and substantial PA phosphatase activity was demonstrated. These data, taken together, supported a phospholipase D-PA phosphatase pathway of DAG formation. Kinetic data established that the low ratio of [^3H]PA/[^3H]DAG formed in reactions with sn-2-[^3H]DOPC was due to a 15-fold higher V"m"a"x and 7-fold lower apparent K"m of the PA phosphatase. The [^3H]PA/[^3H]DAG product ratio was increased by addition of unlabeled PA or by selective extraction of phospholipase D with Triton X-100. The characteristics of the phospholipase D indicated a unique enzyme. Activity was optimal in the presence of EDTA and was almost totally dependent upon Triton X-100. The pH profile displayed a peak at 7.0. Of particular significance was the stringent substrate specificity. Phosphatidylinositol was not hydrolyzed, and activities towards phosphatidylethanolamine and sphingomyelin were at most 30- to 50-fold lower than those towards PC. Phospholipase D and PA phosphatase were identified in a number of rat tissues and other cells. The highest activities of phospholipase D were present in lung and endothelial cells. Phospholipase D was partially purified from rat lung by Triton X-100 extraction and anion exchange chromatography. When linked with PA phosphatase, the phospholipase D could initiate a pathway of DAG formation that is highly specific for PC. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Martin, T.W. oth in Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism Amsterdam : Elsevier 962(1988), 3, Seite 282-296 (DE-627)NLEJ185856497 (DE-600)2209461-1 0005-2760 nnns volume:962 year:1988 number:3 pages:282-296 http://linkinghub.elsevier.com/retrieve/pii/0005-2760(88)90258-5 GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 962 1988 3 282-296 |
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(DE-627)NLEJ185922686 (DE-599)GBVNLZ185922686 DE-627 ger DE-627 rakwb eng Formation of diacylglycerol by a phospholipase D-phosphatidate phosphatase pathway specific for phosphatidylcholine in endothelial cells 1988 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The conversion of phosphatidylcholine (PC) to diacylglycerol (DAG) was studied in sonicated endothelial cells and in subcellular fractions in the presence of 0.05% Triton X-100 and 2 mM EDTA. DAG formation occurred predominantly in an organelle fraction that sedimented at 15 000 x g. In parallel reactions with exogenous 1-oleoyl-2-[^3H]oleoyl-PC (sn-2-[^3H]DOPC) and phosphatidyl[^3H]choline ([choline-^3H]PC), [^3H]DAG was formed by a reaction pathway in which [^3H]choline was the only product derived from [choline-^3H]PC. [^3H]Choline was not formed secondarily from [^3H]glycerophosphocholine or [^3H]phosphocholine. Small amounts of [^3H]phosphatidate ([^3H]PA) were isolated from reactions with sn-2-[^3H]DOPC at short incubation times, and substantial PA phosphatase activity was demonstrated. These data, taken together, supported a phospholipase D-PA phosphatase pathway of DAG formation. Kinetic data established that the low ratio of [^3H]PA/[^3H]DAG formed in reactions with sn-2-[^3H]DOPC was due to a 15-fold higher V"m"a"x and 7-fold lower apparent K"m of the PA phosphatase. The [^3H]PA/[^3H]DAG product ratio was increased by addition of unlabeled PA or by selective extraction of phospholipase D with Triton X-100. The characteristics of the phospholipase D indicated a unique enzyme. Activity was optimal in the presence of EDTA and was almost totally dependent upon Triton X-100. The pH profile displayed a peak at 7.0. Of particular significance was the stringent substrate specificity. Phosphatidylinositol was not hydrolyzed, and activities towards phosphatidylethanolamine and sphingomyelin were at most 30- to 50-fold lower than those towards PC. Phospholipase D and PA phosphatase were identified in a number of rat tissues and other cells. The highest activities of phospholipase D were present in lung and endothelial cells. Phospholipase D was partially purified from rat lung by Triton X-100 extraction and anion exchange chromatography. When linked with PA phosphatase, the phospholipase D could initiate a pathway of DAG formation that is highly specific for PC. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Martin, T.W. oth in Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism Amsterdam : Elsevier 962(1988), 3, Seite 282-296 (DE-627)NLEJ185856497 (DE-600)2209461-1 0005-2760 nnns volume:962 year:1988 number:3 pages:282-296 http://linkinghub.elsevier.com/retrieve/pii/0005-2760(88)90258-5 GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 962 1988 3 282-296 |
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(DE-627)NLEJ185922686 (DE-599)GBVNLZ185922686 DE-627 ger DE-627 rakwb eng Formation of diacylglycerol by a phospholipase D-phosphatidate phosphatase pathway specific for phosphatidylcholine in endothelial cells 1988 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The conversion of phosphatidylcholine (PC) to diacylglycerol (DAG) was studied in sonicated endothelial cells and in subcellular fractions in the presence of 0.05% Triton X-100 and 2 mM EDTA. DAG formation occurred predominantly in an organelle fraction that sedimented at 15 000 x g. In parallel reactions with exogenous 1-oleoyl-2-[^3H]oleoyl-PC (sn-2-[^3H]DOPC) and phosphatidyl[^3H]choline ([choline-^3H]PC), [^3H]DAG was formed by a reaction pathway in which [^3H]choline was the only product derived from [choline-^3H]PC. [^3H]Choline was not formed secondarily from [^3H]glycerophosphocholine or [^3H]phosphocholine. Small amounts of [^3H]phosphatidate ([^3H]PA) were isolated from reactions with sn-2-[^3H]DOPC at short incubation times, and substantial PA phosphatase activity was demonstrated. These data, taken together, supported a phospholipase D-PA phosphatase pathway of DAG formation. Kinetic data established that the low ratio of [^3H]PA/[^3H]DAG formed in reactions with sn-2-[^3H]DOPC was due to a 15-fold higher V"m"a"x and 7-fold lower apparent K"m of the PA phosphatase. The [^3H]PA/[^3H]DAG product ratio was increased by addition of unlabeled PA or by selective extraction of phospholipase D with Triton X-100. The characteristics of the phospholipase D indicated a unique enzyme. Activity was optimal in the presence of EDTA and was almost totally dependent upon Triton X-100. The pH profile displayed a peak at 7.0. Of particular significance was the stringent substrate specificity. Phosphatidylinositol was not hydrolyzed, and activities towards phosphatidylethanolamine and sphingomyelin were at most 30- to 50-fold lower than those towards PC. Phospholipase D and PA phosphatase were identified in a number of rat tissues and other cells. The highest activities of phospholipase D were present in lung and endothelial cells. Phospholipase D was partially purified from rat lung by Triton X-100 extraction and anion exchange chromatography. When linked with PA phosphatase, the phospholipase D could initiate a pathway of DAG formation that is highly specific for PC. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Martin, T.W. oth in Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism Amsterdam : Elsevier 962(1988), 3, Seite 282-296 (DE-627)NLEJ185856497 (DE-600)2209461-1 0005-2760 nnns volume:962 year:1988 number:3 pages:282-296 http://linkinghub.elsevier.com/retrieve/pii/0005-2760(88)90258-5 GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 962 1988 3 282-296 |
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(DE-627)NLEJ185922686 (DE-599)GBVNLZ185922686 DE-627 ger DE-627 rakwb eng Formation of diacylglycerol by a phospholipase D-phosphatidate phosphatase pathway specific for phosphatidylcholine in endothelial cells 1988 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The conversion of phosphatidylcholine (PC) to diacylglycerol (DAG) was studied in sonicated endothelial cells and in subcellular fractions in the presence of 0.05% Triton X-100 and 2 mM EDTA. DAG formation occurred predominantly in an organelle fraction that sedimented at 15 000 x g. In parallel reactions with exogenous 1-oleoyl-2-[^3H]oleoyl-PC (sn-2-[^3H]DOPC) and phosphatidyl[^3H]choline ([choline-^3H]PC), [^3H]DAG was formed by a reaction pathway in which [^3H]choline was the only product derived from [choline-^3H]PC. [^3H]Choline was not formed secondarily from [^3H]glycerophosphocholine or [^3H]phosphocholine. Small amounts of [^3H]phosphatidate ([^3H]PA) were isolated from reactions with sn-2-[^3H]DOPC at short incubation times, and substantial PA phosphatase activity was demonstrated. These data, taken together, supported a phospholipase D-PA phosphatase pathway of DAG formation. Kinetic data established that the low ratio of [^3H]PA/[^3H]DAG formed in reactions with sn-2-[^3H]DOPC was due to a 15-fold higher V"m"a"x and 7-fold lower apparent K"m of the PA phosphatase. The [^3H]PA/[^3H]DAG product ratio was increased by addition of unlabeled PA or by selective extraction of phospholipase D with Triton X-100. The characteristics of the phospholipase D indicated a unique enzyme. Activity was optimal in the presence of EDTA and was almost totally dependent upon Triton X-100. The pH profile displayed a peak at 7.0. Of particular significance was the stringent substrate specificity. Phosphatidylinositol was not hydrolyzed, and activities towards phosphatidylethanolamine and sphingomyelin were at most 30- to 50-fold lower than those towards PC. Phospholipase D and PA phosphatase were identified in a number of rat tissues and other cells. The highest activities of phospholipase D were present in lung and endothelial cells. Phospholipase D was partially purified from rat lung by Triton X-100 extraction and anion exchange chromatography. When linked with PA phosphatase, the phospholipase D could initiate a pathway of DAG formation that is highly specific for PC. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Martin, T.W. oth in Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism Amsterdam : Elsevier 962(1988), 3, Seite 282-296 (DE-627)NLEJ185856497 (DE-600)2209461-1 0005-2760 nnns volume:962 year:1988 number:3 pages:282-296 http://linkinghub.elsevier.com/retrieve/pii/0005-2760(88)90258-5 GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 962 1988 3 282-296 |
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(DE-627)NLEJ185922686 (DE-599)GBVNLZ185922686 DE-627 ger DE-627 rakwb eng Formation of diacylglycerol by a phospholipase D-phosphatidate phosphatase pathway specific for phosphatidylcholine in endothelial cells 1988 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier The conversion of phosphatidylcholine (PC) to diacylglycerol (DAG) was studied in sonicated endothelial cells and in subcellular fractions in the presence of 0.05% Triton X-100 and 2 mM EDTA. DAG formation occurred predominantly in an organelle fraction that sedimented at 15 000 x g. In parallel reactions with exogenous 1-oleoyl-2-[^3H]oleoyl-PC (sn-2-[^3H]DOPC) and phosphatidyl[^3H]choline ([choline-^3H]PC), [^3H]DAG was formed by a reaction pathway in which [^3H]choline was the only product derived from [choline-^3H]PC. [^3H]Choline was not formed secondarily from [^3H]glycerophosphocholine or [^3H]phosphocholine. Small amounts of [^3H]phosphatidate ([^3H]PA) were isolated from reactions with sn-2-[^3H]DOPC at short incubation times, and substantial PA phosphatase activity was demonstrated. These data, taken together, supported a phospholipase D-PA phosphatase pathway of DAG formation. Kinetic data established that the low ratio of [^3H]PA/[^3H]DAG formed in reactions with sn-2-[^3H]DOPC was due to a 15-fold higher V"m"a"x and 7-fold lower apparent K"m of the PA phosphatase. The [^3H]PA/[^3H]DAG product ratio was increased by addition of unlabeled PA or by selective extraction of phospholipase D with Triton X-100. The characteristics of the phospholipase D indicated a unique enzyme. Activity was optimal in the presence of EDTA and was almost totally dependent upon Triton X-100. The pH profile displayed a peak at 7.0. Of particular significance was the stringent substrate specificity. Phosphatidylinositol was not hydrolyzed, and activities towards phosphatidylethanolamine and sphingomyelin were at most 30- to 50-fold lower than those towards PC. Phospholipase D and PA phosphatase were identified in a number of rat tissues and other cells. The highest activities of phospholipase D were present in lung and endothelial cells. Phospholipase D was partially purified from rat lung by Triton X-100 extraction and anion exchange chromatography. When linked with PA phosphatase, the phospholipase D could initiate a pathway of DAG formation that is highly specific for PC. Elsevier Journal Backfiles on ScienceDirect 1907 - 2002 Martin, T.W. oth in Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism Amsterdam : Elsevier 962(1988), 3, Seite 282-296 (DE-627)NLEJ185856497 (DE-600)2209461-1 0005-2760 nnns volume:962 year:1988 number:3 pages:282-296 http://linkinghub.elsevier.com/retrieve/pii/0005-2760(88)90258-5 GBV_USEFLAG_H ZDB-1-SDJ GBV_NL_ARTICLE AR 962 1988 3 282-296 |
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Formation of diacylglycerol by a phospholipase D-phosphatidate phosphatase pathway specific for phosphatidylcholine in endothelial cells |
| abstract |
The conversion of phosphatidylcholine (PC) to diacylglycerol (DAG) was studied in sonicated endothelial cells and in subcellular fractions in the presence of 0.05% Triton X-100 and 2 mM EDTA. DAG formation occurred predominantly in an organelle fraction that sedimented at 15 000 x g. In parallel reactions with exogenous 1-oleoyl-2-[^3H]oleoyl-PC (sn-2-[^3H]DOPC) and phosphatidyl[^3H]choline ([choline-^3H]PC), [^3H]DAG was formed by a reaction pathway in which [^3H]choline was the only product derived from [choline-^3H]PC. [^3H]Choline was not formed secondarily from [^3H]glycerophosphocholine or [^3H]phosphocholine. Small amounts of [^3H]phosphatidate ([^3H]PA) were isolated from reactions with sn-2-[^3H]DOPC at short incubation times, and substantial PA phosphatase activity was demonstrated. These data, taken together, supported a phospholipase D-PA phosphatase pathway of DAG formation. Kinetic data established that the low ratio of [^3H]PA/[^3H]DAG formed in reactions with sn-2-[^3H]DOPC was due to a 15-fold higher V"m"a"x and 7-fold lower apparent K"m of the PA phosphatase. The [^3H]PA/[^3H]DAG product ratio was increased by addition of unlabeled PA or by selective extraction of phospholipase D with Triton X-100. The characteristics of the phospholipase D indicated a unique enzyme. Activity was optimal in the presence of EDTA and was almost totally dependent upon Triton X-100. The pH profile displayed a peak at 7.0. Of particular significance was the stringent substrate specificity. Phosphatidylinositol was not hydrolyzed, and activities towards phosphatidylethanolamine and sphingomyelin were at most 30- to 50-fold lower than those towards PC. Phospholipase D and PA phosphatase were identified in a number of rat tissues and other cells. The highest activities of phospholipase D were present in lung and endothelial cells. Phospholipase D was partially purified from rat lung by Triton X-100 extraction and anion exchange chromatography. When linked with PA phosphatase, the phospholipase D could initiate a pathway of DAG formation that is highly specific for PC. |
| abstractGer |
The conversion of phosphatidylcholine (PC) to diacylglycerol (DAG) was studied in sonicated endothelial cells and in subcellular fractions in the presence of 0.05% Triton X-100 and 2 mM EDTA. DAG formation occurred predominantly in an organelle fraction that sedimented at 15 000 x g. In parallel reactions with exogenous 1-oleoyl-2-[^3H]oleoyl-PC (sn-2-[^3H]DOPC) and phosphatidyl[^3H]choline ([choline-^3H]PC), [^3H]DAG was formed by a reaction pathway in which [^3H]choline was the only product derived from [choline-^3H]PC. [^3H]Choline was not formed secondarily from [^3H]glycerophosphocholine or [^3H]phosphocholine. Small amounts of [^3H]phosphatidate ([^3H]PA) were isolated from reactions with sn-2-[^3H]DOPC at short incubation times, and substantial PA phosphatase activity was demonstrated. These data, taken together, supported a phospholipase D-PA phosphatase pathway of DAG formation. Kinetic data established that the low ratio of [^3H]PA/[^3H]DAG formed in reactions with sn-2-[^3H]DOPC was due to a 15-fold higher V"m"a"x and 7-fold lower apparent K"m of the PA phosphatase. The [^3H]PA/[^3H]DAG product ratio was increased by addition of unlabeled PA or by selective extraction of phospholipase D with Triton X-100. The characteristics of the phospholipase D indicated a unique enzyme. Activity was optimal in the presence of EDTA and was almost totally dependent upon Triton X-100. The pH profile displayed a peak at 7.0. Of particular significance was the stringent substrate specificity. Phosphatidylinositol was not hydrolyzed, and activities towards phosphatidylethanolamine and sphingomyelin were at most 30- to 50-fold lower than those towards PC. Phospholipase D and PA phosphatase were identified in a number of rat tissues and other cells. The highest activities of phospholipase D were present in lung and endothelial cells. Phospholipase D was partially purified from rat lung by Triton X-100 extraction and anion exchange chromatography. When linked with PA phosphatase, the phospholipase D could initiate a pathway of DAG formation that is highly specific for PC. |
| abstract_unstemmed |
The conversion of phosphatidylcholine (PC) to diacylglycerol (DAG) was studied in sonicated endothelial cells and in subcellular fractions in the presence of 0.05% Triton X-100 and 2 mM EDTA. DAG formation occurred predominantly in an organelle fraction that sedimented at 15 000 x g. In parallel reactions with exogenous 1-oleoyl-2-[^3H]oleoyl-PC (sn-2-[^3H]DOPC) and phosphatidyl[^3H]choline ([choline-^3H]PC), [^3H]DAG was formed by a reaction pathway in which [^3H]choline was the only product derived from [choline-^3H]PC. [^3H]Choline was not formed secondarily from [^3H]glycerophosphocholine or [^3H]phosphocholine. Small amounts of [^3H]phosphatidate ([^3H]PA) were isolated from reactions with sn-2-[^3H]DOPC at short incubation times, and substantial PA phosphatase activity was demonstrated. These data, taken together, supported a phospholipase D-PA phosphatase pathway of DAG formation. Kinetic data established that the low ratio of [^3H]PA/[^3H]DAG formed in reactions with sn-2-[^3H]DOPC was due to a 15-fold higher V"m"a"x and 7-fold lower apparent K"m of the PA phosphatase. The [^3H]PA/[^3H]DAG product ratio was increased by addition of unlabeled PA or by selective extraction of phospholipase D with Triton X-100. The characteristics of the phospholipase D indicated a unique enzyme. Activity was optimal in the presence of EDTA and was almost totally dependent upon Triton X-100. The pH profile displayed a peak at 7.0. Of particular significance was the stringent substrate specificity. Phosphatidylinositol was not hydrolyzed, and activities towards phosphatidylethanolamine and sphingomyelin were at most 30- to 50-fold lower than those towards PC. Phospholipase D and PA phosphatase were identified in a number of rat tissues and other cells. The highest activities of phospholipase D were present in lung and endothelial cells. Phospholipase D was partially purified from rat lung by Triton X-100 extraction and anion exchange chromatography. When linked with PA phosphatase, the phospholipase D could initiate a pathway of DAG formation that is highly specific for PC. |
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Formation of diacylglycerol by a phospholipase D-phosphatidate phosphatase pathway specific for phosphatidylcholine in endothelial cells |
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http://linkinghub.elsevier.com/retrieve/pii/0005-2760(88)90258-5 |
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7.399349 |