title: Post-translational modifications modulate ligand recognition by the third PDZ domain of the MAGUK protein PSD-95.
abstract: The relative promiscuity of hub proteins such as postsynaptic density protein-95 (PSD-95) can be achieved by alternative splicing, allosteric regulation, and post-translational modifications, the latter of which is the most efficient method of accelerating cellular responses to environmental changes in vivo. Here, a mutational approach was used to determine the impact of phosphorylation and succinimidation post-translational modifications on the binding affinity of the postsynaptic density protein-95/discs large/zonula occludens-1 (PDZ3) domain of PSD-95. Molecular dynamics simulations revealed that the binding affinity of this domain is influenced by an interplay between salt-bridges linking the α3 helix, the β2-β3 loop and the positively charged Lys residues in its high-affinity hexapeptide ligand KKETAV. The α3 helix is an extra structural element that is not present in other PDZ domains, which links PDZ3 with the following SH3 domain in the PSD-95 protein. This regulatory mechanism was confirmed experimentally via thermodynamic and NMR chemical shift perturbation analyses, discarding intra-domain long-range effects. Taken together, the results presented here reveal the molecular basis of the regulatory role of the α3 extra-element and the effects of post-translational modifications of PDZ3 on its binding affinity, both energetically and dynamically.
authors.affiliation: Department of Physical Chemistry and Institute of Biotechnology, Faculty of Sciences, University of Granada, Granada, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Murciano-Calles
authors.affiliation: Department of Physical Chemistry and Institute of Biotechnology, Faculty of Sciences, University of Granada, Granada, Spain.
authors.forename: Carles
authors.initials: C
authors.lastname: Corbi-Verge
authors.affiliation: Department of Physical Chemistry and Institute of Biotechnology, Faculty of Sciences, University of Granada, Granada, Spain.
authors.forename: Adela M
authors.initials: AM
authors.lastname: Candel
authors.affiliation: Department of Physical Chemistry and Institute of Biotechnology, Faculty of Sciences, University of Granada, Granada, Spain.
authors.forename: Irene
authors.initials: I
authors.lastname: Luque
authors.affiliation: Department of Physical Chemistry and Institute of Biotechnology, Faculty of Sciences, University of Granada, Granada, Spain.
authors.forename: Jose C
authors.initials: JC
authors.lastname: Martinez
mesh_terms: D000577:Amides / Q000378:metabolism; D000595:Amino Acid Sequence; D008024:Ligands; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D054731:PDZ Domains; D010766:Phosphorylation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017433:Protein Structure, Secondary; D055672:Static Electricity; D013386:Succinates / Q000378:metabolism; D013816:Thermodynamics
chemical_list: D000577:Amides; D008024:Ligands; D008565:Membrane Proteins; D013386:Succinates; C011116:succinamide