The talin rod IBS2 alpha-helix interacts with the beta3 integrin cytoplasmic tail membrane-proximal helix by establishing charge complementary salt bridges.

Talin establishes a major link between integrins and actin filaments and contains two distinct integrin binding sites: one, IBS1, located in the talin head domain and involved in integrin activation and a second, IBS2, that maps to helix 50 of the talin rod domain and is essential for linking integrin β subunits to the cytoskeleton (Moes, M., Rodius, S., Coleman, S. J., Monkley, S. J., Goormaghtigh, E., Tremuth, L., Kox, C., van der Holst, P. P., Critchley, D. R., and Kieffer, N. (2007) J. Biol. Chem. 282,17280 -17288). Through the combined approach of mutational analysis of the β3 integrin cytoplasmic tail and the talin rod IBS2 site, SPR binding studies, as well as site-specific antibody inhibition experiments, we provide evidence that the integrin β3-talin rod interaction relies on a helix-helix association between α-helix 50 of the talin rod domain and the membrane-proximal α-helix of the β3 integrin cytoplasmic tail. Moreover, charge complementarity between the highly conserved talin rod IBS2 lysine residues and integrin β3 glutamic acid residues is necessary for this interaction. Our results support a model in which talin IBS2 binds to the same face of the β3 subunit cytoplasmic helix as the integrin αIIb cytoplasmic tail helix, suggesting that IBS2 can only interact with the β3 subunit following integrin activation.