| Chapter title |
Direct and Continuous Measurement of Phospholipase D Activities Using the Chelation-Enhanced Fluorescence Property of 8-Hydroxyquinoline
|
|---|---|
| Chapter number | 6 |
| Book title |
Lipases and Phospholipases
|
| Published in |
Methods in molecular biology, August 2018
|
| DOI | 10.1007/978-1-4939-8672-9_6 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-8671-2, 978-1-4939-8672-9
|
| Authors |
Renaud Rahier, Houda Abla, Yani Arhab, Alexandre Noiriel, Abdelkarim Abousalham, Rahier, Renaud, Abla, Houda, Arhab, Yani, Noiriel, Alexandre, Abousalham, Abdelkarim |
| Abstract |
Phospholipase D (PLD) hydrolyzes phospholipids to form phosphatidic acid (PA) and the corresponding headgroup. To date, PLD has been linked to several pathologies, such as cancer, making this enzyme an important therapeutic target. However, most PLD assays developed so far are either discontinuous or based on the indirect determination of choline released upon phosphatidylcholine (PC) hydrolysis. Therefore, we designed a PLD assay that is based on the chelation-enhanced fluorescence property of 8-hydroxyquinoline. This assay exhibits a strong fluorescence signal upon Ca2+ complexation with the PLD-generated PA and is not limited to PC as the substrate but allows the use of natural phospholipids with various headgroups. Besides, this easy-to-handle assay allows to monitor prokaryotic and eukaryotic PLD activities in a continuous way and on a microplate scale. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 4 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 2 | 50% |
| Professor | 1 | 25% |
| Unknown | 1 | 25% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 1 | 25% |
| Neuroscience | 1 | 25% |
| Unknown | 2 | 50% |