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Discovery, Structure–Activity Relationship Studies, and Crystal Structure of Nonpeptide Inhibitors Bound to the Shank3 PDZ Domain
Author(s) -
Saupe Jörn,
Roske Yvette,
Schillinger Christian,
Kamdem Nestor,
Radetzki Silke,
Diehl Anne,
Oschkinat Hartmut,
Krause Gerd,
Heinemann Udo,
Rademann Jörg
Publication year - 2011
Publication title -
chemmedchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.817
H-Index - 100
eISSN - 1860-7187
pISSN - 1860-7179
DOI - 10.1002/cmdc.201100094
Subject(s) - pdz domain , postsynaptic density , heteronuclear single quantum coherence spectroscopy , scaffold protein , allosteric regulation , guanylate kinase , biology , biophysics , chemistry , microbiology and biotechnology , biochemistry , nuclear magnetic resonance spectroscopy , postsynaptic potential , stereochemistry , signal transduction , receptor , membrane protein , membrane
Shank is the central scaffolding protein of the postsynaptic density (PSD) protein complex found in cells of the central nervous system. Cellular studies indicate a prominent role of the protein in the organization of the PSD, in the development of neuronal morphology, in neuronal signaling, and in synaptic plasticity, thus linking Shank functions to the molecular basis of learning and memory. Mutations in the Shank gene have been found in several neuronal disorders including mental retardation, typical autism, and Asperger syndrome. Shank is linked to the PSD complex via its PDZ domain that binds to the C‐terminus of guanylate‐kinase‐associated protein (GKAP). Here, small‐molecule inhibitors of Shank3 PDZ domain are developed. A fluorescence polarization assay based on an identified high‐affinity peptide is established, and tetrahydroquinoline carboxylates are identified as inhibitors of this protein–protein interaction. Chemical synthesis via a hetero‐Diels–Alder strategy is employed for hit optimization, and structure–activity relationship studies are performed. Best hits possess K i values in the 10 μ M range, and binding to the PDZ domain is confirmed by 1 H, 15 N HSQC NMR experiments. One of the hits crystallizes with the Shank3 PDZ domain. The structure, analyzed at a resolution of 1.85 Å, reveals details of the binding mode. Finally, binding to PDZ domains of PSD‐95, syntrophin, and DVL3 was studied using 1 H, 15 N HSQC NMR spectroscopy.