The neuronal Ca2+-binding protein 2 (NECAB2) interacts with the adenosine A2A receptor and modulates the cell surface expression and function of the receptor

Canela, Laia; Luján, Rafael; Lluı́s, Carme; Burgueño, Javier; Mallol, Josefa; Canela, Enric I.; Franco, Rafael; Ciruela, Francisco

doi:10.1016/j.mcn.2007.05.007

Cited by 41 publications

(54 citation statements)

References 36 publications

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“…The function of NECAB2 (Ca 2+ buffer or sensor) at the spinal level is not known, but studies in some brain regions may give a lead. Thus, in rat hippocampus and hypothalamus, studies on mGluR5 and α2a-adrenergic receptor suggest a sensor role (22,26). NECAB2 was identified as the interacting protein for the nuclear receptor coactivator vitamin D receptor-interacting protein complex component 150 (59), which also supports a "sensor"-like function for NECAB2 (60).…”

Section: Discussionmentioning

confidence: 98%

“…NECABs are also EF-hand proteins, with one pair of EF-hand motifs in the N terminus and a putative antibiotic biosynthesis monooxygenase domain in the C terminus, which are linked by a NECAB homogeneous region (22). NECAB1/2 are restricted to the nervous system, whereas NECAB3 is also expressed in the heart and skeletal muscle (21).…”

mentioning

confidence: 99%

“…It encodes a 389-aa (NECAB2) (24). NECAB2 was identified as a downstream target of Pax6 in mouse retina, which is involved in retinal development (24,25), as well as being a binding partner for the adenosine A 2A receptor (22). Furthermore, an interaction between NECAB2 and metabotropic glutamate receptor 5 (mGluR5) was demonstrated in rat hippocampal pyramidal cells, possibly regulating mGluR5's coupling to its signaling machinery (26).…”

mentioning

confidence: 99%

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Neuronal calcium-binding proteins 1/2 localize to dorsal root ganglia and excitatory spinal neurons and are regulated by nerve injury

Zhang¹,

Tortoriello

Hsueh

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Neuronal calcium (Ca 2+ )-binding proteins 1 and 2 (NECAB1/2) are members of the phylogenetically conserved EF-hand Ca 2+ -binding protein superfamily. To date, NECABs have been explored only to a limited extent and, so far, not at all at the spinal level. Here, we describe the distribution, phenotype, and nerve injury-induced regulation of NECAB1/NECAB2 in mouse dorsal root ganglia (DRGs) and spinal cord. In DRGs, NECAB1/2 are expressed in around 70% of mainly small-and medium-sized neurons. Many colocalize with calcitonin gene-related peptide and isolectin B4, and thus represent nociceptors. NECAB1/2 neurons are much more abundant in DRGs than the Ca 2+ -binding proteins (parvalbumin, calbindin, calretinin, and secretagogin) studied to date. In the spinal cord, the NECAB1/2 distribution is mainly complementary. NECAB1 labels interneurons and a plexus of processes in superficial layers of the dorsal horn, commissural neurons in the intermediate area, and motor neurons in the ventral horn. Using CLARITY, a novel, bilaterally connected neuronal system with dendrites that embrace the dorsal columns like palisades is observed. NECAB2 is present in cell bodies and presynaptic boutons across the spinal cord. In the dorsal horn, most NECAB1/2 neurons are glutamatergic. Both NECAB1/2 are transported into dorsal roots and peripheral nerves. Peripheral nerve injury reduces NECAB2, but not NECAB1, expression in DRG neurons. Our study identifies NECAB1/2 as abundant Ca 2+ -binding proteins in painrelated DRG neurons and a variety of spinal systems, providing molecular markers for known and unknown neuron populations of mechanosensory and pain circuits in the spinal cord.commissural interneurons | GAD67 | neuropathic pain | VGLUT2

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Section: Discussionmentioning

confidence: 98%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Neuronal calcium-binding proteins 1/2 localize to dorsal root ganglia and excitatory spinal neurons and are regulated by nerve injury

Zhang¹,

Tortoriello

Hsueh

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…A2A-ct interacts with a number of proteins, including G-protein-coupled receptor kinases (GRKs), b-arrestins, a-actinins, calmodulin (CaM), neuronal calcium-binding protein 2 (NECAB2), translin-associated protein X (TRAX), Arf nucleotide-binding site opener (ARNO/cytohesin-2), ubiquitin-specific protease 4 (USP4), and neuronal calcium sensor protein 1 (NCS-1) (17)(18)(19)(20)(21)(22)(23)(24). These interactions have been studied using yeast two-hybrid screens, colocalization, coimmunoprecipitation, pulldown, immunoelectron microscopy, mass spectrometry, and bioluminescence and fluorescence resonance energy transfer approaches using either crude brain membrane extracts containing A2AR, A2AR-transfected cell cultures, or small synthetic A2AR peptides.…”

Section: The Human Adenosine A2a Receptormentioning

confidence: 99%

Human Adenosine A2A Receptor Binds Calmodulin with High Affinity in a Calcium-Dependent Manner

Piirainen

Hellman

Tossavainen

et al. 2015

Biophysical Journal

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Understanding how ligands bind to G-protein-coupled receptors and how binding changes receptor structure to affect signaling is critical for developing a complete picture of the signal transduction process. The adenosine A2A receptor (A2AR) is a particularly interesting example, as it has an exceptionally long intracellular carboxyl terminus, which is predicted to be mainly disordered. Experimental data on the structure of the A2AR C-terminus is lacking, because published structures of A2AR do not include the C-terminus. Calmodulin has been reported to bind to the A2AR C-terminus, with a possible binding site on helix 8, next to the membrane. The biological meaning of the interaction as well as its calcium dependence, thermodynamic parameters, and organization of the proteins in the complex are unclear. Here, we characterized the structure of the A2AR C-terminus and the A2AR C-terminus-calmodulin complex using different biophysical methods, including native gel and analytical gel filtration, isothermal titration calorimetry, NMR spectroscopy, and small-angle X-ray scattering. We found that the C-terminus is disordered and flexible, and it binds with high affinity (Kd = 98 nM) to calmodulin without major conformational changes in the domain. Calmodulin binds to helix 8 of the A2AR in a calcium-dependent manner that can displace binding of A2AR to lipid vesicles. We also predicted and classified putative calmodulin-binding sites in a larger group of G-protein-coupled receptors.

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“…It has been suggested that the length of A 2A -ct and the lack of membrane anchoring render the domain less constrained and more flexible, enabling A 2A -ct-accessory protein interactions and A 2A R subsequent signal transmission through G-protein-independent pathways (Keuerleber et al 2011). Several proteins interacting with A 2A -ct have been identified, including other signalling proteins such as ARNO/cytohesin-2 (Gsandtner et al 2005), elementary in MAPK activation, NECAB2 (Canela et al 2007), which enhances A 2A R intracellular retention and signalling through MAPK, TRAX (Sun et al 2006) involved in regulation of neurite growth, and calmodulin which modulates the function of A 2A R-dopamine D 2 receptor complexes (Woods et al 2008).…”

Section: Biological Contextmentioning

confidence: 99%

HN, N, Cα, Cβ and C′ assignments of the intrinsically disordered C-terminus of human adenosine A2A receptor

et al. 2015

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The C-terminus of the human adenosine A2A receptor differs from the other human adenosine receptors by its exceptional length and lack of a canonical cysteine residue. We have previously structurally characterized this C-terminal domain and its interaction with calmodulin. It was shown to be structurally disordered and flexible, and to bind calmodulin with high affinity in a calcium-dependent manner. Interaction with calmodulin takes place at the N-terminal end of the A2A C-terminal domain without major conformational changes in the latter. NMR was one of the biophysical methods used in the study. Here we present the H(N), N, C(α), C(β) and C' chemical shift assignments of the free form of the C-terminus residues 293-412, used in the NMR spectroscopic characterization of the domain.

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The neuronal Ca2+-binding protein 2 (NECAB2) interacts with the adenosine A2A receptor and modulates the cell surface expression and function of the receptor

Cited by 41 publications

References 36 publications

Neuronal calcium-binding proteins 1/2 localize to dorsal root ganglia and excitatory spinal neurons and are regulated by nerve injury

Neuronal calcium-binding proteins 1/2 localize to dorsal root ganglia and excitatory spinal neurons and are regulated by nerve injury

Human Adenosine A2A Receptor Binds Calmodulin with High Affinity in a Calcium-Dependent Manner

HN, N, Cα, Cβ and C′ assignments of the intrinsically disordered C-terminus of human adenosine A2A receptor

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