title: Gelsolin binding to phosphatidylinositol 4,5-bisphosphate is modulated by calcium and pH.
abstract: The actin cytoskeleton of nonmuscle cells undergoes extensive remodeling during agonist stimulation. Lamellipodial extension is initiated by uncapping of actin nuclei at the cortical cytoplasm to allow filament elongation. Many actin filament capping proteins are regulated by phosphatidylinositol 4,5-bisphosphate (PIP2), which is hydrolyzed by phospholipase C. It is hypothesized that PIP2 dissociates capping proteins from filament ends to promote actin assembly. However, since actin polymerization often occurs at a time when PIP2 concentration is decreased rather than increased, capping protein interactions with PIP2 may not be regulated solely by the bulk PIP2 concentration. We present evidence that PIP2 binding to the gelsolin family of capping proteins is enhanced by Ca2+. Binding was examined by equilibrium and nonequilibrium gel filtration and by monitoring intrinsic tryptophan fluorescence. Gelsolin and CapG affinity for PIP2 were increased 8- and 4-fold, respectively, by microM Ca2+, and the Ca2+ requirement was reduced by lowering the pH from 7.5 to 7.0. Studies with the NH2- and COOH-terminal halves of gelsolin showed that PIP2 binding occurred primarily at the NH2-terminal half, and Ca2+ exposed its PIP2 binding sites through a change in the COOH-terminal half. Mild acidification promotes PIP2 binding by directly affecting the NH2-terminal sites. Our findings can explain increased PIP2-induced uncapping even as the PIP2 concentration drops during cell activation. The change in gelsolin family PIP2 binding affinity during cell activation can impact divergent PIP2-dependent processes by altering PIP2 availability. Cross-talk between these proteins provides a multilayered mechanism for positive and negative modulation of signal transduction from the plasma membrane to the cytoskeleton.
authors.affiliation: Department of Physiology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: K M
authors.initials: KM
authors.lastname: Lin
authors.forename: E
authors.initials: E
authors.lastname: Wenegieme
authors.forename: P J
authors.initials: PJ
authors.lastname: Lu
authors.forename: C S
authors.initials: CS
authors.lastname: Chen
authors.forename: H L
authors.initials: HL
authors.lastname: Yin
mesh_terms: D002118:Calcium / Q000502:physiology*; D004533:Egtazic Acid / Q000494:pharmacology; D018260:Gelsolin / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008840:Microfilament Proteins / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*
chemical_list: D018260:Gelsolin; D008840:Microfilament Proteins; D009687:Nuclear Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D004533:Egtazic Acid; D002118:Calcium