Selectivity and promiscuity of the first and second PDZ domains of PSD-95 and synapse-associated protein 102

J Biol Chem. 2002 Jun 14;277(24):21697-711. doi: 10.1074/jbc.M112339200. Epub 2002 Apr 5.

Abstract

PDZ domains typically interact with the very carboxyl terminus of their binding partners. Type 1 PDZ domains usually require valine, leucine, or isoleucine at the very COOH-terminal (P(0)) position, and serine or threonine 2 residues upstream at P(-2). We quantitatively defined the contributions of carboxyl-terminal residues to binding selectivity of the prototypic interactions of the PDZ domains of postsynaptic density protein 95 (PSD-95) and its homolog synapse-associated protein 90 (SAP102) with the NR2b subunit of the N-methyl-d-aspartate-type glutamate receptor. Our studies indicate that all of the last five residues of NR2b contribute to the binding selectivity. Prominent were a requirement for glutamate or glutamine at P(-3) and for valine at P(0) for high affinity binding and a preference for threonine over serine at P(-2), in the context of the last 11 residues of the NR2b COOH terminus. This analysis predicts a COOH-terminal (E/Q)(S/T)XV consensus sequence for the strongest binding to the first two PDZ domains of PSD-95 and SAP102. A search of the human genome sequences for proteins with a COOH-terminal (E/Q)(S/T)XV motif yielded 50 proteins, many of which have not been previously identified as PSD-95 or SAP102 binding partners. Two of these proteins, brain-specific angiogenesis inhibitor 1 and protein kinase Calpha, co-immunoprecipitated with PSD-95 and SAP102 from rat brain extracts.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Angiogenesis Inhibitors
  • Angiogenic Proteins*
  • Animals
  • Anisotropy
  • Brain / metabolism
  • Disks Large Homolog 4 Protein
  • Dose-Response Relationship, Drug
  • Frizzled Receptors
  • Genome, Human
  • Glutathione Transferase / metabolism
  • Humans
  • Immunoblotting
  • Intracellular Signaling Peptides and Proteins
  • Isoenzymes / chemistry
  • Kinetics
  • Membrane Proteins
  • Models, Molecular
  • Molecular Sequence Data
  • Nerve Tissue Proteins / chemistry*
  • Neuropeptides / chemistry*
  • Nuclear Proteins*
  • Peptides / chemistry
  • Precipitin Tests
  • Protein Binding
  • Protein Kinase C / chemistry
  • Protein Kinase C-alpha
  • Protein Structure, Tertiary
  • Proteins / chemistry
  • Rats
  • Receptors, G-Protein-Coupled
  • Receptors, Neurotransmitter / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Serine / chemistry
  • Spectrometry, Fluorescence
  • Threonine / chemistry
  • Transcription Factors*

Substances

  • ADGRB1 protein, human
  • Angiogenesis Inhibitors
  • Angiogenic Proteins
  • DLG3 protein, human
  • Disks Large Homolog 4 Protein
  • Dlg3 protein, rat
  • Dlg4 protein, rat
  • FZD2 protein, human
  • Frizzled Receptors
  • Intracellular Signaling Peptides and Proteins
  • Isoenzymes
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Neuropeptides
  • Nuclear Proteins
  • Peptides
  • Proteins
  • Receptors, G-Protein-Coupled
  • Receptors, Neurotransmitter
  • Recombinant Fusion Proteins
  • Transcription Factors
  • postsynaptic density proteins
  • Fzd2 protein, rat
  • Threonine
  • Serine
  • Glutathione Transferase
  • PRKCA protein, human
  • Protein Kinase C
  • Protein Kinase C-alpha