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Date
10/12/2011 17:56:02
Group number
PI name
Pérez, I.
Channel family
Glutamate-gated sodium and calcium channel (GluR)
Channel name
NMDA-type glutamate receptor (NMDAR)
Channel subunit
NR3A
UNIPROT
Rat NR3A Q9R1M7, Human NR3A Q8TCU5
Relevant publication (1)
Cloning:
Ciabarra et al., Cloning and characterization of chi-1: a developmentally regulated member of a novel class of the ionotropic glutamate receptor family. J Neurosci. 1995; 15(10):6498-508.
Sucher et al., Developmental and regional expression pattern of a novel NMDA receptor-like subunit (NMDAR-L) in the rodent brain. J Neurosci. 1995, 15(10):6509-20.
Relevant publication (2)
Biophysical properties:
Pérez-Otaño et al., Assembly with the NR1 subunit is required for surface expression of NR3A-containing NMDA receptors. J Neurosci. 2001, 21(4):1228-37.
Sasaki et al., Characterization and comparison of the NR3A subunit of the NMDA receptor in recombinant systems and primary cortical neurons. J Neurophysiol. 2002 Apr;87(4):2052-63.
Burzomato et al., The receptor subunits generating NMDA receptor mediated currents in oligodendrocytes. J Physiol. 2010;588(Pt 18):3403-14.
Relevant publication (3)
Function and localization:
Das et al., Increased NMDA current and spine density in mice lacking the NMDA receptor subunit NR3A. Nature. 1998;393(6683):377-81.
Roberts et al., Downregulation of NR3A-containing NMDARs is required for synapse maturation and memory consolidation. Neuron. 2009;63(3):342-56.
Nakanishi et al., Neuroprotection by the NR3A subunit of the NMDA receptor. J Neurosci. 2009, 29(16):5260-5.
Larsen et al., NR3A-containing NMDARs promote neurotransmitter release and spike timing-dependent plasticity. Nat Neurosci. 2011
Henson et al., Influence of the NR3A subunit on NMDA receptor functions. Prog Neurobiol. 2010;91(1):23-37.
Wong et al., Temporal and regional expression of NMDA receptor subunit NR3A in the mammalian brain. J Comp Neurol. 2002; 450(4):303-17.
Wada et al., NR3A modulates the outer vestibule of the "NMDA" receptor channel. J Neurosci. 2006;26(51):13156-66.
CD
FTIR
NMR
Absorbance /
Fluorescence
Interaction protein (1)
PP2A, protein phosphatase 2A (Chan S.F. and Sucher N.J., An NMDA receptor signaling complex with protein phosphatase 2A. J Neurosci. 2001 Oct 15;21(20):7985-92).

PACSIN/syndapin1 (Pérez-Otaño I. et al., Endocytosis and synaptic removal of NR3A-containing NMDA receptors by PACSIN1/syndapin1. Nat Neurosci. 2006 May;9(5):611-21). MAP1S, microtubule associated-protein 1S (Eriksson M. et al., The NMDAR subunit NR3A interacts with microtubule-associated protein 1S in the brain. Biochem Biophys Res Commun. 2007 Sep 14;361(1):127-32).

Plectin (Eriksson M. et al., On the role of NR3A in human NMDA receptors. Physiol Behav. 2007 Sep 10;92(1-2):54-9). CARP1, cell cycle and apoptosis regulatory protein-1 (Eriksson M. et al., On the role of NR3A in human NMDA receptors. Physiol Behav. 2007 Sep 10;92(1-2):54-9).

GPS2, G-protein pathway suppressor 2 (Eriksson M. et al., On the role of NR3A in human NMDA receptors. Physiol Behav. 2007 Sep 10;92(1-2):54-9).

Dynein light chain LC8 (Navarro-Lérida I. et al., Proteomic identification of brain proteins that interact with dynein light chain LC8. Proteomics. 2004 Feb;4(2):339-46).

Rheb, Small GTPase Ras homologue enriched in brain (Sucher N. J. et al., Association of the small GTPase Rheb with the NMDA receptor subunit NR3A. Neurosignals. 2010;18(4):203-9).
Interaction protein (2)
Interaction protein (3)
Interaction protein (4)
PDB (1)
PDB (2)
PDB (3)
Expression tissues
Early postnatal expression in most brain regions (peak around P5-P25), largely down-regulated in adult brain (Henson et al., Influence of the NR3A subunit on NMDA receptor functions. Prog Neurobiol. 2010, 91(1):23-37, Wong et al., Temporal and regional expression of NMDA receptor subunit NR3A in the mammalian brain. J Comp Neurol. 2002, 450(4):303-17).

Expression in human fetal brain (Mueller et al., Expression of the NR3A subunit of the NMDA receptor in human fetal brain. Ann N Y Acad Sci. 2003;1003:448-51)

Expression in oligodendrocytes (Karadottir et al NMDA receptors are expressed in oligodendrocytes and activated in ischaemia. Nature. 2005;438(7071):1162-6; Micu et al., NMDA receptors mediate calcium accumulation in myelin during chemical ischaemia, Nature. 2006; 439(7079):988-92).
Expression cell lines
Inner medullary collecting duct kidney cells (IMCD)
Expression plasmids
Mammalian expression

1. Rat NR3A in pcINeo (Perez-Otaño I et al., Assembly with the NR1 subunit is required for surface expression of NR3A-containing NMDA receptors, J Neurosci (2001), vol. 21, 1228-1237.)

2. Rat NR3A in gWIZ

3. Rat NR3A in lentiviral vector SWS-GFP

4. GFP-NR3A in pcINeo (Perez-Otaño I et al., Endocytosis and synaptic removal of NR3A-containing NMDA receptors by PACSIN1/synadapinI, Nat Neurosci (2006), vol. 9, 611-621.)

5. GFP-NR3A in gWIZ

6. GFP-NR3A in pRK5

7. GFP-NR3A in Sidnbid virus vector pSinRep5

8. HA-NR3A in pcINeo

9. superecliptic GFP-NR3A in pcINeo

10. Tac-NR3A(C-term tail) in pCDM8

Bacterial expression

11. GST-NR3A (C-term tail) in PGEX-4T

12. GST-NR3A (C-term tail) in PGEX-6P-1

Yeast expression

13. Rat NR3A in pAS2-1 (Perez-Otaño I et al., Endocytosis and synaptic removal of NR3A-containing NMDA receptors by PACSIN1/synadapinI, Nat Neurosci (2006), vol. 9, 611-621.)
Level of expression
Channel quaternary
Tetramer (NR1, NR2x, NR3A)
Agonists
Smile agonist
Antagonists and blockers
Smile antagonist
Structure-function
relationship
Molecular determinant assembly
Molecular determinant clustering
Molecular determinant trafficking
Binding to NR1 is required for functional assembly, ER export and plasma membrane expression (Pérez-Otaño et al., Assembly with the NR1 subunit is required for surface expression of NR3A-containing NMDA receptors. J Neurosci. 2001;21(4):1228-37)

Binding to PACSIN/syndapin 1 is required for endocytosis (Pérez-Otaño et al., Endocytosis and synaptic removal of NR3A-containing NMDA receptors by PACSIN1/syndapin1. Nat Neurosci. 2006;9(5):611-21).

Two endocytic signals are contained within the proximal 25 residues of the NR3A carboxy-terminal tail.
Molecular determinant binding
Extracellular ligand binding pocket formed by the S1 (N511 to R660) and S2 segments (E776 to K915)

Awobuluyi et al., Subunit-specific roles of glycine-binding domains in activation of NR1/NR3 N-methyl-D-aspartate receptors. Mol Pharmacol. 2007
Mutants constructs
1. Rat GFPNR3A C-term truncation in pcINeo

2. Rat GFPNR3A C-term truncation (Y695F) in pcINeo

3. Rat GFPNR3A -977 del in pcINeo

4. Rat GFPNR3A -977 del (YWL-AAA) in pcINeo

5. Tac-NR3A-977 del in pCDM8

6. Tac-NR3A-977 del (YWL-AAA) in pCDM8

7. Tac-NR3A-977 del (LL-AA, YWL-AAA) in pCDM8

8. Tac-NR3A-977 del (KSK-AAA) in pCDM8

9. Tac-NR3A-977 del (LQY-AAA) in pCDM8

10. Tac-NR3A-977 del (HTS-AAA) in pCDM8

11. Tac-NR3A-966 del in pCDM8

12. Tac-NR3A-966 del (LL-AA) in pCDM8

13. Tac-NR3A-996 del in pCDM8

14. Tac-NR3A-1003 del in pCDM8

15. Tac-NR3A-1097 del in pCDM8

16. Tac-NR3A-977-1115 in pCDM8

B. Full-length mutants

17. Rat NR3A-Y971F in pcINeo

18. Rat NR3A-Y1028F in pcINeo

19. Rat NR3A-Y1105F in pcINeo

20. HA-NR3A-Y971F in pcINeo

21. HA-NR3A-Y1028F in pcINeo

22. HA-NR3A-Y1105F in pcINeo

23. GFP-NR3A-YWL-AAA (971-973) in pcINeo

24. GFP-NR3A-LMVW-AMVA (1012-1015) in pcINeo

25. GFP-NR3A-Y971E in pcINeo

26. GFP-NR3A-Y971A in pcINeo

27. GFP-NR3A-Y695F in pcINeo

28. GFP-NR3A-Y971F in pcINeo

29. GFP-NR3A-Y1028F in pcINeo

30. GFP-NR3A-Y1105F in pcINeo

31. GFP-NR3A-Y971F/Y1028F/Y1105F in pcINeo

32. NR3A-Y971F/Y1028F/Y1105F in pcINeo

33. HA-NR3A-Y971F/Y1028F/Y1105F in pcINeo

Bacterial expression

A. Truncated mutants

34. GST-NR3A (C-term tail) 977 del in PGEX-4T

35. Mouse GST-NR3A (C-term tail) 1040-1115 in PGEX-4T

36. GST-NR3A (C-term tail) 996 del in PGEX-4T

37. GST-NR3A (C-term tail) 976-1115 in PGEX-4T

38. GST-NR3A (C-term tail) 1031-1115 in PGEX-4T

39. GST-NR3A (C-term tail) 1081-1115 in PGEX-4T

B. Full-length mutants

40. GST-NR3A (C-term tail)- YWL-AAA (971-973) in PGEX-4T

41. GST-NR3A (C-term tail)- WL-AA (972-973) in PGEX-4T

42. GST-NR3A (C-term tail)- Y971E in PGEX-4T

43. GST-NR3A (C-term tail)- Y971A in PGEX-4T

44. GST-NR3A (C-term tail)- Y971F in PGEX-4T

45. GST-NR3A (C-term tail)- L958A,L959A in PGEX-4T

46. GST-NR3A (C-term tail)- YWL-AAA (971-973)/L958A,L959A in PGEX-4T

47. GST-NR3A (C-term tail)- LMVW-AMVA (1012-1015) in PGEX

48. GST-NR3A (C-term tail)-Y971F in PGEX-6P-1

49. GST-NR3A (C-term tail)-3Y-3F in PGEX-6P-1

Yeast expression

A. Truncated mutants

50. Rat NR3A -996 del in pAS2-1

51. Rat NR3A -976-1115 in pAS2-1

52. Rat NR3A -969-1115 in pAS2-1

53. Rat NR3A -1034 del in pAS2-1
Antibodies
NR3A (Millipore, #07-356), Larsen RS et al., NR3A-containing NMDARs promote neurotransmitter release and spike-timing dependent plasticity, Nat Neurosci (2011), vol. 14, 338-344.

NR3A (P-20) (SantaCruz, # sc-51160)

NR3A (Y-20) (SantaCruz, # sc-51162)

Rabbit anti-NR3A DC-1 (generated by our group)
Diseases

 

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