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- Title
Phosphatidylinositol 4,5-bisphosphate (PIP<sub>2</sub>) facilitates norepinephrine transporter dimerization and modulates substrate efflux.
- Authors
Luethi, Dino; Maier, Julian; Rudin, Deborah; Szöllősi, Dániel; Angenoorth, Thomas J. F.; Stankovic, Stevan; Schittmayer, Matthias; Burger, Isabella; Yang, Jae-Won; Jaentsch, Kathrin; Holy, Marion; Das, Anand Kant; Brameshuber, Mario; Camacho-Hernandez, Gisela Andrea; Casiraghi, Andrea; Newman, Amy Hauck; Kudlacek, Oliver; Birner-Gruenberger, Ruth; Stockner, Thomas; Schütz, Gerhard J.
- Abstract
The plasmalemmal norepinephrine transporter (NET) regulates cardiovascular sympathetic activity by clearing extracellular norepinephrine in the synaptic cleft. Here, we investigate the subunit stoichiometry and function of NET using single-molecule fluorescence microscopy and flux assays. In particular, we show the effect of phosphatidylinositol 4,5-bisphosphate (PIP2) on NET oligomerization and efflux. NET forms monomers (~60%) and dimers (~40%) at the plasma membrane. PIP2 depletion results in a decrease in the average oligomeric state and decreases NET-mediated substrate efflux while not affecting substrate uptake. Mutation of the putative PIP2 binding residues R121, K334, and R440 to alanines does not affect NET dimerization but results in decreased substrate efflux that is not altered upon PIP2 depletion; this indicates that PIP2 interactions with these residues affect NET-mediated efflux. A dysregulation of norepinephrine and PIP2 signaling have both been implicated in neuropsychiatric and cardiovascular diseases. This study provides evidence that PIP2 directly regulates NET organization and function. Phosphatidylinositol 4,5-bisphosphate (PIP2) directly regulates norepinephrine transporter (NET) dimer stabilization at the plasma membrane. Furthermore, PIP2 interaction with NET affects efflux.
- Subjects
NORADRENALINE; DIMERIZATION; CELL membranes; FLUORESCENCE microscopy; P-glycoprotein; DIMERS
- Publication
Communications Biology, 2022, Vol 5, Issue 1, p1
- ISSN
2399-3642
- Publication type
Article
- DOI
10.1038/s42003-022-04210-1