Lian, Lu-Yun, Pandalaneni, Sravan R, Patel, Pryank, McCue, Hannah V, Haynes, Lee P ORCID: 0000-0002-1296-0338 and Burgoyne, Robert D ORCID: 0000-0002-9219-0387
(2011)
Characterisation of the Interaction of the C-Terminus of the Dopamine D2 Receptor with Neuronal Calcium Sensor-1.
PLOS ONE, 6 (11).
e27779-.
Text
Characterisation of the interaction of the C-terminus of the dopamine D2 receptor with neuronal calcium sensor-1.pdf - Published version Download (1MB) | Preview |
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.
Item Type: | Article |
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Uncontrolled Keywords: | Animals, Humans, Rats, Neuropeptides, Peptide Fragments, Receptors, Dopamine D2, Magnetic Resonance Spectroscopy, Calcium Signaling, Amino Acid Sequence, Protein Structure, Tertiary, Protein Binding, Models, Molecular, Molecular Sequence Data, Neuronal Calcium-Sensor Proteins, Protein Multimerization |
Depositing User: | Symplectic Admin |
Date Deposited: | 11 Nov 2019 10:57 |
Last Modified: | 19 Jan 2023 00:20 |
DOI: | 10.1371/journal.pone.0027779 |
Related URLs: | |
URI: | https://livrepository.liverpool.ac.uk/id/eprint/3061304 |