Characterisation of the interaction of the C-terminus of the dopamine D2 receptor with neuronal calcium sensor-1

Lu-Yun Lian, Sravan R. Pandalaneni, Pryank Patel, Hannah V. McCue, Lee P. Haynes, Robert D. Burgoyne

Research output: Contribution to journalArticlepeer-review

27 Citations (Scopus)
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Abstract

NCS-1 is a member of the neuronal calcium sensor (NCS) family of EF-hand Ca(2+) binding proteins which has been implicated in several physiological functions including regulation of neurotransmitter release, membrane traffic, voltage gated Ca(2+) channels, neuronal development, synaptic plasticity, and learning. NCS-1 binds to the dopamine D2 receptor, potentially affecting its internalisation and controlling dopamine D2 receptor surface expression. The D2 receptor binds NCS-1 via a short 16-residue cytoplasmic C-terminal tail. We have used NMR and fluorescence spectroscopy to characterise the interactions between the NCS-1/Ca(2+) and D2 peptide. The data show that NCS-1 binds D2 peptide with a K(d) of ∼14.3 µM and stoichiometry of peptide binding to NCS-1 of 2:1. NMR chemical shift mapping confirms that D2 peptide binds to the large, solvent-exposed hydrophobic groove, on one face of the NCS-1 molecule, with residues affected by the presence of the peptide spanning both the N and C-terminal portions of the protein. The NMR and mutagenesis data further show that movement of the C-terminal helix 11 of NCS-1 to fully expose the hydrophobic groove is important for D2 peptide binding. Molecular docking using restraints derived from the NMR chemical shift data, together with the experimentally-derived stoichiometry, produced a model of the complex between NCS-1 and the dopamine receptor, in which two molecules of the receptor are able to simultaneously bind to the NCS-1 monomer.

Original languageEnglish
Pages (from-to)e27779
JournalPLoS ONE
Volume6
Issue number11
DOIs
Publication statusPublished - 16 Nov 2011

Keywords

  • Amino Acid Sequence
  • Animals
  • Calcium Signaling
  • Humans
  • Magnetic Resonance Spectroscopy
  • Models, Molecular
  • Molecular Sequence Data
  • Neuronal Calcium-Sensor Proteins
  • Neuropeptides
  • Peptide Fragments
  • Protein Binding
  • Protein Multimerization
  • Protein Structure, Tertiary
  • Rats
  • Receptors, Dopamine D2

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