Formation of nNOS/PSD-95 PDZ dimer requires a preformed beta-finger structure from the nNOS PDZ domain

PDZ domains are modular protein units that play important roles in organizi ng signal transduction complexes. PDZ domains mediate interactions with bot h C-terminal peptide ligands and other PDZ domains. Here, we used PDZ domai ns from neuronal nitric oxide synthase (nNOS) and postsynaptic density prot ein-95 (PSD-95) to explore the mechanism for PDZ-dimer formation. The nNOS PDZ domain terminates with a similar to 30 residue amino acid beta -finger peptide that is shown to be required for nNOS/PSD-95 PDZ dimer formation. L n addition, formation of the PDZ dimer requires this beta -finger peptide t o be physically anchored to the main body of the canonical nNOS PDZ domain. A buried salt bridge between the beta -finger and the PDZ domain induces a nd stabilizes the beta -hairpin structure of the nNOS PDZ domain. In apo-nN OS, the beta -finger peptide is partially flexible and adopts a transient b eta -strand like structure that is stabilized upon PDZ dimer formation. The flexibility of the NOS PDZ beta -finger is likely to play a critical role in supporting the formation of nNOS/PSD-95 complex. The experimental data a lso suggest that nNOS PDZ and the second PDZ domain of PSD-95 form a "head- to-tail" dimer similar to the nNOS/syntrophin complex characterized by X-ra y crystallography. (C) 2000 Academic Press.