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Fluorogenic XY-69 in Lipid Vesicles for Measuring Activity of Phospholipase C Isozymes.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2251, pp. 225-236. - Publication Year :
- 2021
-
Abstract
- Mammalian phospholipase C (PLC) isozymes are major signaling nodes that regulate a wide range of cellular processes. Dysregulation of PLC activity has been associated with a growing list of human diseases such as cancer and Alzheimer's disease. However, methods to directly and continuously monitor PLC activity at membranes with high sensitivity and throughput are still lacking. We have developed XY-69, a fluorogenic PIP <subscript>2</subscript> analog, which can be efficiently hydrolyzed by PLC isozymes either in solution or at membranes. Here, we describe the optimized assay conditions and protocol to measure the activity of PLC-γ1 (D1165H) with XY-69 in lipid vesicles. The described protocol also applies to other PLC isozymes.
- Subjects :
- Fluorescein-5-isothiocyanate chemistry
Hydrolysis
Isoenzymes analysis
Lipid Metabolism physiology
Lipids chemistry
Phosphatidylinositol 4,5-Diphosphate chemistry
Phospholipase C gamma analysis
Phospholipase C gamma metabolism
Protein Binding physiology
Type C Phospholipases chemistry
Type C Phospholipases metabolism
Enzyme Assays methods
Phosphatidylinositol 4,5-Diphosphate analogs & derivatives
Type C Phospholipases analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 2251
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 33481244
- Full Text :
- https://doi.org/10.1007/978-1-0716-1142-5_17