Protein kinase C (PKC) plays a critical role in diseases such as
cancer,
stroke, and cardiac
ischemia, and participates in a variety of signal transduction pathways such as apoptosis, cell proliferation, and
tumor suppression. Though much is known about PKC downstream signaling events, the mechanisms of regulation of PKC activation and subsequent translocation have not been elucidated.
Protein-
protein interactions regulate and determine the specificity of many cellular signaling events. Such a specific
protein-
protein interaction is described here between deltaPKC and
annexin V. We demonstrate, at physiologically relevant conditions, that a transient interaction between
annexin V and deltaPKC occurs in cells after deltaPKC stimulation, but before deltaPKC translocates to the particulate fraction. Evidence of deltaPKC-
annexin V binding is provided also by FRET and by in vitro binding studies. Dissociation of the deltaPKC-
annexin V complex requires
ATP and microtubule integrity. Furthermore, depletion of endogenous
annexin V, but not
annexin IV, with
siRNA inhibits deltaPKC translocation following PKC stimulation. A rationally designed eight
amino acid peptide, corresponding to the interaction site for deltaPKC on
annexin V, inhibits deltaPKC translocation and deltaPKC-mediated function as evidenced by its protective effect in a model of
myocardial infarction. Our data indicate that translocation of deltaPKC is not simply a diffusion-driven process, but is instead a multi-step event regulated by
protein-
protein interactions. We show that following cell activation, deltaPKC-
annexin V binding is a transient and an essential step in the function of deltaPKC, thus identifying a new role for
annexin V in PKC signaling and a new step in PKC activation.