Back to Search
Start Over
Multistep Compositional Remodeling of Supported Lipid Membranes by Interfacially Active Phosphatidylinositol Kinases
- Source :
- Analytical chemistry, vol 88, iss 10
- Publication Year :
- 2016
- Publisher :
- eScholarship, University of California, 2016.
-
Abstract
- The multienzyme catalytic phosphorylation of phosphatidylinositol (PI) in a supported lipid membrane platform is demonstrated for the first time. One-step treatment with PI 4-kinase IIIβ (PI4Kβ) yielded PI 4-phosphate (PI4P), while a multistep enzymatic cascade of PI4Kβ followed by PIP 5-kinase produced PI-4,5-bisphosphate (PI(4,5)P2 or PIP2). By employing quartz crystal microbalance with dissipation monitoring, we were able to track membrane association of kinase enzymes for the first time as well as detect PI4P and PI(4,5)P2 generation based on subsequent antibody binding to the supported lipid bilayers. Pharmacologic inhibition of PI4Kβ by a small molecule inhibitor was also quantitatively assessed, yielding an EC50 value that agrees well with conventional biochemical readout. Taken together, the development of a PI-containing supported membrane platform coupled with surface-sensitive measurement techniques for kinase studies opens the door to exploring the rich biochemistry and pharmacological targeting of membrane-associated phosphoinositides.
- Subjects :
- Lipid Bilayers
Membrane Proteins
Analytical Chemistry
Phosphotransferases (Alcohol Group Acceptor)
Phosphatidylinositol 4
Phosphatidylinositol Phosphates
5.1 Pharmaceuticals
5-Diphosphate
Quartz Crystal Microbalance Techniques
Generic health relevance
Enzyme Inhibitors
Development of treatments and therapeutic interventions
Other Chemical Sciences
1-Phosphatidylinositol 4-Kinase
Subjects
Details
- Database :
- OpenAIRE
- Journal :
- Analytical chemistry, vol 88, iss 10
- Accession number :
- edsair.od.......325..20daebced97e3976782c5b65fba39293