Characterization of Lysophospholipase D Activity in Mammalian Cell Membranes
Lysophosphatidic acid (LPA) is a lipid mediator that binds to G-protein-coupled receptors, eliciting a wide variety of responses in mammalian cells. Lyso-phospholipids generated via phospholipase A<sub>2</sub> (PLA<sub>2</sub>) can be converted to LPA by a lysophospholipase D...
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MDPI AG
2024-03-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/13/6/520 |
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| author | Yuhuan Xie Krishna M. Ella Terra C. Gibbs Marianne E. Yohannan Stewart M. Knoepp Pravita Balijepalli G. Patrick Meier Kathryn E. Meier |
| author_facet | Yuhuan Xie Krishna M. Ella Terra C. Gibbs Marianne E. Yohannan Stewart M. Knoepp Pravita Balijepalli G. Patrick Meier Kathryn E. Meier |
| author_sort | Yuhuan Xie |
| collection | DOAJ |
| description | Lysophosphatidic acid (LPA) is a lipid mediator that binds to G-protein-coupled receptors, eliciting a wide variety of responses in mammalian cells. Lyso-phospholipids generated via phospholipase A<sub>2</sub> (PLA<sub>2</sub>) can be converted to LPA by a lysophospholipase D (lyso-PLD). Secreted lyso-PLDs have been studied in more detail than membrane-localized lyso-PLDs. This study utilized in vitro enzyme assays with fluorescent substrates to examine LPA generation in membranes from multiple mammalian cell lines (PC12, rat pheochromocytoma; A7r5, rat vascular smooth muscle; Rat-1, rat fibroblast; PC-3, human prostate carcinoma; and SKOV-3 and OVCAR-3, human ovarian carcinoma). The results show that membranes contain a lyso-PLD activity that generates LPA from a fluorescent alkyl-lyso-phosphatidylcholine, as well as from naturally occurring acyl-linked lysophospholipids. Membrane lyso-PLD and PLD activities were distinguished by multiple criteria, including lack of effect of PLD2 over-expression on lyso-PLD activity and differential sensitivities to vanadate (PLD inhibitor) and iodate (lyso-PLD inhibitor). Based on several lines of evidence, including siRNA knockdown, membrane lyso-PLD is distinct from autotaxin, a secreted lyso-PLD. PC-3 cells express GDE4 and GDE7, recently described lyso-PLDs that localize to membranes. These findings demonstrate that membrane-associated lyso-D activity, expressed by multiple mammalian cell lines, can contribute to LPA production. |
| first_indexed | 2024-04-24T18:26:57Z |
| format | Article |
| id | doaj.art-7d4978fdcd01418ab0d3880c7c45722f |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-04-24T18:26:57Z |
| publishDate | 2024-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-7d4978fdcd01418ab0d3880c7c45722f2024-03-27T13:30:39ZengMDPI AGCells2073-44092024-03-0113652010.3390/cells13060520Characterization of Lysophospholipase D Activity in Mammalian Cell MembranesYuhuan Xie0Krishna M. Ella1Terra C. Gibbs2Marianne E. Yohannan3Stewart M. Knoepp4Pravita Balijepalli5G. Patrick Meier6Kathryn E. Meier7Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Pharmaceutical Sciences, College of Pharmacy and Pharmaceutical Sciences, Washington State University, Spokane, WA 99202, USADepartment of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Pharmaceutical Sciences, College of Pharmacy and Pharmaceutical Sciences, Washington State University, Spokane, WA 99202, USALysophosphatidic acid (LPA) is a lipid mediator that binds to G-protein-coupled receptors, eliciting a wide variety of responses in mammalian cells. Lyso-phospholipids generated via phospholipase A<sub>2</sub> (PLA<sub>2</sub>) can be converted to LPA by a lysophospholipase D (lyso-PLD). Secreted lyso-PLDs have been studied in more detail than membrane-localized lyso-PLDs. This study utilized in vitro enzyme assays with fluorescent substrates to examine LPA generation in membranes from multiple mammalian cell lines (PC12, rat pheochromocytoma; A7r5, rat vascular smooth muscle; Rat-1, rat fibroblast; PC-3, human prostate carcinoma; and SKOV-3 and OVCAR-3, human ovarian carcinoma). The results show that membranes contain a lyso-PLD activity that generates LPA from a fluorescent alkyl-lyso-phosphatidylcholine, as well as from naturally occurring acyl-linked lysophospholipids. Membrane lyso-PLD and PLD activities were distinguished by multiple criteria, including lack of effect of PLD2 over-expression on lyso-PLD activity and differential sensitivities to vanadate (PLD inhibitor) and iodate (lyso-PLD inhibitor). Based on several lines of evidence, including siRNA knockdown, membrane lyso-PLD is distinct from autotaxin, a secreted lyso-PLD. PC-3 cells express GDE4 and GDE7, recently described lyso-PLDs that localize to membranes. These findings demonstrate that membrane-associated lyso-D activity, expressed by multiple mammalian cell lines, can contribute to LPA production.https://www.mdpi.com/2073-4409/13/6/520lipid mediatorsphospholipasessignal transduction |
| spellingShingle | Yuhuan Xie Krishna M. Ella Terra C. Gibbs Marianne E. Yohannan Stewart M. Knoepp Pravita Balijepalli G. Patrick Meier Kathryn E. Meier Characterization of Lysophospholipase D Activity in Mammalian Cell Membranes Cells lipid mediators phospholipases signal transduction |
| title | Characterization of Lysophospholipase D Activity in Mammalian Cell Membranes |
| title_full | Characterization of Lysophospholipase D Activity in Mammalian Cell Membranes |
| title_fullStr | Characterization of Lysophospholipase D Activity in Mammalian Cell Membranes |
| title_full_unstemmed | Characterization of Lysophospholipase D Activity in Mammalian Cell Membranes |
| title_short | Characterization of Lysophospholipase D Activity in Mammalian Cell Membranes |
| title_sort | characterization of lysophospholipase d activity in mammalian cell membranes |
| topic | lipid mediators phospholipases signal transduction |
| url | https://www.mdpi.com/2073-4409/13/6/520 |
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