GTPase activating proteins (GAPs) of the centaurin family regulate the actin cytoskeleton and vesicle trafficking through inactivation of the ADP-ribosylation factor (ARF) family of small GTP-binding proteins (1-5). We report the functional characterisation of centaurin-α2, which is structurally related to centaurin-α1 ARF6 GAP (2). Centaurin-α2 contains an N-terminal GAP domain followed by two pleckstrin homology (PH) domains (N-PH and C-PH). In vitro, GFP-centaurin-α2 specifically binds phosphatidylinositol (PI) 3-kinase lipid products, PI 3,4-P2 and PI 3,4,5-P3 (PIP3), through the C-terminal PH domain. In agreement with this observation, GFP-centaurin-α2 was recruited to the plasma membrane from the cytosol in EGF-stimulated cells in a PI 3-kinase-dependent manner (2). Moreover, the C-PH domain is sufficient and necessary for membrane recruitment of centaurin-α2. Centaurin-α2 shows sustained kinetics of PI 3-kinase-mediated membrane recruitment in EGF-stimulated cells, due to its binding to PI 3,4-P2 (2). Centaurin-α2 prevents ARF6 translocation to and cortical actin formation at the plasma membrane, which are phenotypic indications for ARF6 activation, in EGF-stimulated cells. Moreover, the constitutively active mutant of ARF6 reverses the effect of centaurin-α2 on cortical actin formation. The membrane targeted centaurin-α2 is constitutively active. Together, these studies indicate that centaurin-α2 recruits in a sustained manner to the plasma membrane through binding to PI 3,4-P2 and thereby regulates actin reorganisation via ARF6 (2).