Ci-VSP is a depolarization-activated phosphatidylinositol-4,5-bisphosphate and phosphatidylinositol-3,4,5-trisphosphate 5'-phosphatase.
Phosphoinositides are membrane-delimited regulators of protein function and control many different cellular targets. The differentially phosphorylated isoforms have distinct concentrations in various subcellular membranes, which can change dynamically in response to cellular signaling events. Maintenance and dynamics of phosphoinositide levels involve a complex set of enzymes, among them phospholipases and lipid kinases and phosphatases. Recently, a novel type of phosphoinositide-converting protein (termed Ci-VSP) that contains a voltage sensor domain was isolated. It was already shown that Ci-VSP can alter phosphoinositide levels in a voltage-dependent manner. However, the exact enzymatic reaction catalyzed by Ci-VSP is not known. We used fluorescent phosphoinositide-binding probes and total internal reflection microscopy together with patch-clamp measurements from living cells to delineate substrates and products of Ci-VSP. Upon activation of Ci-VSP by membrane depolarization, membrane association of phosphatidylinositol (PI) (4,5)P2- and PI(3,4,5)P3-specific binding domains decreased, revealing consumption of these phosphoinositides by Ci-VSP. Depletion of PI(4,5)P2 was coincident with an increase in membrane PI(4)P. Similarly, PI(3,4)P2 was generated during depletion of PI(3,4,5)P3. These results suggest that Ci-VSP acts as a 5'-phosphatase of PI(4,5)P2 and PI(3,4,5)P3.
Halaszovich
Christian R
CR
Department of Neurophysiology, Institute of Physiology and Pathophysiology, Philipps-Universität Marburg, 35037 Marburg, Germany.
Schreiber
Daniela N
DN
Oliver
Dominik
D
D000818:Animals; D016466:CHO Cells; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D004594:Electrophysiology; D004789:Enzyme Activation; D018408:Patch-Clamp Techniques; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism
D019269:Phosphatidylinositol 4,5-Diphosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C096024:phosphatidylinositol-3,4,5-trisphosphate 5-phosphatase; D010744:Phosphoric Monoester Hydrolases