Lysosomal glycerophosphocholine phosphodiesterase in Tetrahymena
The purification and characterization of a novel phosphodiesterase (PDE) is presented. The activity was detected in the extracellular medium of Tetrahymena thermophila cultures, by the release of p-nitrophenol from p-nitrophenylphosphocholine (PNPPC) with an acidic pH optimum. In cell homogenates, i...
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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_10399712_v47_n2_p283_FlorinChristensen |
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todo:paper_10399712_v47_n2_p283_FlorinChristensen2023-10-03T15:57:38Z Lysosomal glycerophosphocholine phosphodiesterase in Tetrahymena Florin-Christensen, J. Florin-Christensen, M. Extracellular hydrolase Glycerophosphocholine phosphodiesterase Lipid metabolism Lysosomal Tetrahymena 4 nitrophenol acid phosphatase choline (4 nitrophenyl phosphate) glycerol glycerophosphocholine phosphodiesterase glycerophosphorylcholine hydrolase phosphatidylcholine unclassified drug article cell free system culture medium degradation enzyme activity enzyme analysis enzyme purification enzyme substrate latent period lysosome nonhuman parasite cultivation pH polyacrylamide gel electrophoresis tetrahymena thermophila Animals Enzyme Stability Glycerylphosphorylcholine Hydrogen-Ion Concentration Kinetics Lysosomes Phosphatidylcholines Phosphoric Diester Hydrolases Phosphorylcholine Substrate Specificity Temperature Tetrahymena Eukaryota Protozoa Tetrahymena Tetrahymena thermophila The purification and characterization of a novel phosphodiesterase (PDE) is presented. The activity was detected in the extracellular medium of Tetrahymena thermophila cultures, by the release of p-nitrophenol from p-nitrophenylphosphocholine (PNPPC) with an acidic pH optimum. In cell homogenates, it is sedimentable, shows a latency similar to that of acid phosphatase and is co-secreted with this enzyme, indicating that it is a lysosomal hydrolase. PNPPC-PDE was purified to homogeneity from the extracellular medium, yielding a single band of 58 kD on SDS polyacrylamide gel electrophoresis. It catalyzed the release of glycerol from glycerophosphocholine (GPC) and GPC competitively inhibits degradation of PNPPC. We present further evidence indicating that the natural substrate for PNPPC-PDE is GPC. Thus, Tetrahymena becomes the first eukaryote in which a lysosomal GPC-PDE is observed. This finding provides a new pathway for the complete breakdown of phosphatidylcholine in a lysosomal medium. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_10399712_v47_n2_p283_FlorinChristensen |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Extracellular hydrolase Glycerophosphocholine phosphodiesterase Lipid metabolism Lysosomal Tetrahymena 4 nitrophenol acid phosphatase choline (4 nitrophenyl phosphate) glycerol glycerophosphocholine phosphodiesterase glycerophosphorylcholine hydrolase phosphatidylcholine unclassified drug article cell free system culture medium degradation enzyme activity enzyme analysis enzyme purification enzyme substrate latent period lysosome nonhuman parasite cultivation pH polyacrylamide gel electrophoresis tetrahymena thermophila Animals Enzyme Stability Glycerylphosphorylcholine Hydrogen-Ion Concentration Kinetics Lysosomes Phosphatidylcholines Phosphoric Diester Hydrolases Phosphorylcholine Substrate Specificity Temperature Tetrahymena Eukaryota Protozoa Tetrahymena Tetrahymena thermophila |
| spellingShingle |
Extracellular hydrolase Glycerophosphocholine phosphodiesterase Lipid metabolism Lysosomal Tetrahymena 4 nitrophenol acid phosphatase choline (4 nitrophenyl phosphate) glycerol glycerophosphocholine phosphodiesterase glycerophosphorylcholine hydrolase phosphatidylcholine unclassified drug article cell free system culture medium degradation enzyme activity enzyme analysis enzyme purification enzyme substrate latent period lysosome nonhuman parasite cultivation pH polyacrylamide gel electrophoresis tetrahymena thermophila Animals Enzyme Stability Glycerylphosphorylcholine Hydrogen-Ion Concentration Kinetics Lysosomes Phosphatidylcholines Phosphoric Diester Hydrolases Phosphorylcholine Substrate Specificity Temperature Tetrahymena Eukaryota Protozoa Tetrahymena Tetrahymena thermophila Florin-Christensen, J. Florin-Christensen, M. Lysosomal glycerophosphocholine phosphodiesterase in Tetrahymena |
| topic_facet |
Extracellular hydrolase Glycerophosphocholine phosphodiesterase Lipid metabolism Lysosomal Tetrahymena 4 nitrophenol acid phosphatase choline (4 nitrophenyl phosphate) glycerol glycerophosphocholine phosphodiesterase glycerophosphorylcholine hydrolase phosphatidylcholine unclassified drug article cell free system culture medium degradation enzyme activity enzyme analysis enzyme purification enzyme substrate latent period lysosome nonhuman parasite cultivation pH polyacrylamide gel electrophoresis tetrahymena thermophila Animals Enzyme Stability Glycerylphosphorylcholine Hydrogen-Ion Concentration Kinetics Lysosomes Phosphatidylcholines Phosphoric Diester Hydrolases Phosphorylcholine Substrate Specificity Temperature Tetrahymena Eukaryota Protozoa Tetrahymena Tetrahymena thermophila |
| description |
The purification and characterization of a novel phosphodiesterase (PDE) is presented. The activity was detected in the extracellular medium of Tetrahymena thermophila cultures, by the release of p-nitrophenol from p-nitrophenylphosphocholine (PNPPC) with an acidic pH optimum. In cell homogenates, it is sedimentable, shows a latency similar to that of acid phosphatase and is co-secreted with this enzyme, indicating that it is a lysosomal hydrolase. PNPPC-PDE was purified to homogeneity from the extracellular medium, yielding a single band of 58 kD on SDS polyacrylamide gel electrophoresis. It catalyzed the release of glycerol from glycerophosphocholine (GPC) and GPC competitively inhibits degradation of PNPPC. We present further evidence indicating that the natural substrate for PNPPC-PDE is GPC. Thus, Tetrahymena becomes the first eukaryote in which a lysosomal GPC-PDE is observed. This finding provides a new pathway for the complete breakdown of phosphatidylcholine in a lysosomal medium. |
| format |
JOUR |
| author |
Florin-Christensen, J. Florin-Christensen, M. |
| author_facet |
Florin-Christensen, J. Florin-Christensen, M. |
| author_sort |
Florin-Christensen, J. |
| title |
Lysosomal glycerophosphocholine phosphodiesterase in Tetrahymena |
| title_short |
Lysosomal glycerophosphocholine phosphodiesterase in Tetrahymena |
| title_full |
Lysosomal glycerophosphocholine phosphodiesterase in Tetrahymena |
| title_fullStr |
Lysosomal glycerophosphocholine phosphodiesterase in Tetrahymena |
| title_full_unstemmed |
Lysosomal glycerophosphocholine phosphodiesterase in Tetrahymena |
| title_sort |
lysosomal glycerophosphocholine phosphodiesterase in tetrahymena |
| url |
http://hdl.handle.net/20.500.12110/paper_10399712_v47_n2_p283_FlorinChristensen |
| work_keys_str_mv |
AT florinchristensenj lysosomalglycerophosphocholinephosphodiesteraseintetrahymena AT florinchristensenm lysosomalglycerophosphocholinephosphodiesteraseintetrahymena |
| _version_ |
1807323895743643648 |