cGMP-Phosphodiesterase 6 (PDE6) is the central effector enzyme in the phototransduction system of vertebrate photoreceptors. We have recently found that PDE6 accumulates in a detergent-resistant membrane (DRM) fraction in response to excitation of bovine rod phototransduction system. Here, we studied the molecular mechanism of the PDE6 translocation to DRM. Pertussis toxin inhibited the translocation of PDE6. Upon addition of AlF4- to dark-adapted ROS, PDE6 translocated to DRM along with a minor fraction of the alpha-subunit of transducin (Talpha). The addition of an excess of the inhibitory subunit of PDE6 blocked its accumulation in the DRM, but did not block the translocation of the minor fraction of Talpha. These data suggested that the formation of a complex between activated Talpha and PDE6 imparted upon Talpha a high affinity for the DRM. The translocation of PDE6 to the DRM may be involved in the spatiotemporal regulation of its activity on disk membranes. (C) 2003 Elsevier Science (USA). All rights reserved.