Thermodynamics and structural analysis of positive allosteric modulation of the ionotropic glutamate receptor GluA2
(2012) In Biochemical Journal 441(1). p.173-178- Abstract
- Positive allosteric modulators of the ionotropic glutamate receptor-2 (GluA2) are promising compounds for the treatment of cognitive disorders, e.g. Alzheimer's disease. These modulators bind within the dimer interface of the LBD (ligand-binding domain) and stabilize the agonist-bound conformation slowing receptor desensitization and/or deactivation. In the present study, we employ isothermal titration calorimetry to determine binding affinities and thermodynamic details of binding of modulators of GluA2. A mutant of the LBD of GluA2 (LBD-L483Y-N754S) that forms a stable dimer in solution was used. The potent GluA2 modulator BPAM-97 was used as a reference compound. Evidence that BPAM-97 binds in the same pocket as the well-known GluA2... (More)
- Positive allosteric modulators of the ionotropic glutamate receptor-2 (GluA2) are promising compounds for the treatment of cognitive disorders, e.g. Alzheimer's disease. These modulators bind within the dimer interface of the LBD (ligand-binding domain) and stabilize the agonist-bound conformation slowing receptor desensitization and/or deactivation. In the present study, we employ isothermal titration calorimetry to determine binding affinities and thermodynamic details of binding of modulators of GluA2. A mutant of the LBD of GluA2 (LBD-L483Y-N754S) that forms a stable dimer in solution was used. The potent GluA2 modulator BPAM-97 was used as a reference compound. Evidence that BPAM-97 binds in the same pocket as the well-known GluA2 modulator cyclothiazide was obtained from X-ray structures. The LBD-L483Y-N754S:BPAM-97 complex has a Kd of 5.6 μM (ΔH=-4.9 kcal/mol, -TΔS=-2.3 kcal/mol; where 1 kcal≈4.187 kJ). BPAM-97 was used in a displacement assay to determine a Kd of 0.46 mM (ΔH=-1.2 kcal/mol, -TΔS=-3.3 kcal/mol) for the LBD-L483Y-N754S:IDRA-21 complex. The major structural factors increasing the potency of BPAM-97 over IDRA-21 are the increased van der Waals contacts to, primarily, Met496 in GluA2 imposed by the ethyl substituent of BPAM-97. These results add important information on binding affinities and thermodynamic details, and provide a new tool in the development of drugs against cognitive disorders. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/aa7928dd-476a-4b8f-bfef-80ce4547781c
- author
- Krintel, Christian LU ; Frydenvang, Karla ; Olsen, Lars ; Kristensen, Maria T ; de Barrios, Oriol ; Naur, Peter ; Francotte, Pierre ; Pirotte, Bernard ; Gajhede, Michael and Kastrup, Jette S
- publishing date
- 2012
- type
- Contribution to journal
- publication status
- published
- in
- Biochemical Journal
- volume
- 441
- issue
- 1
- pages
- 173 - 178
- publisher
- Portland Press
- external identifiers
-
- scopus:84055219498
- ISSN
- 0264-6021
- DOI
- 10.1042/BJ20111221
- language
- English
- LU publication?
- no
- id
- aa7928dd-476a-4b8f-bfef-80ce4547781c
- date added to LUP
- 2017-06-10 06:51:34
- date last changed
- 2022-06-21 14:25:38
@article{aa7928dd-476a-4b8f-bfef-80ce4547781c, abstract = {{Positive allosteric modulators of the ionotropic glutamate receptor-2 (GluA2) are promising compounds for the treatment of cognitive disorders, e.g. Alzheimer's disease. These modulators bind within the dimer interface of the LBD (ligand-binding domain) and stabilize the agonist-bound conformation slowing receptor desensitization and/or deactivation. In the present study, we employ isothermal titration calorimetry to determine binding affinities and thermodynamic details of binding of modulators of GluA2. A mutant of the LBD of GluA2 (LBD-L483Y-N754S) that forms a stable dimer in solution was used. The potent GluA2 modulator BPAM-97 was used as a reference compound. Evidence that BPAM-97 binds in the same pocket as the well-known GluA2 modulator cyclothiazide was obtained from X-ray structures. The LBD-L483Y-N754S:BPAM-97 complex has a Kd of 5.6 μM (ΔH=-4.9 kcal/mol, -TΔS=-2.3 kcal/mol; where 1 kcal≈4.187 kJ). BPAM-97 was used in a displacement assay to determine a Kd of 0.46 mM (ΔH=-1.2 kcal/mol, -TΔS=-3.3 kcal/mol) for the LBD-L483Y-N754S:IDRA-21 complex. The major structural factors increasing the potency of BPAM-97 over IDRA-21 are the increased van der Waals contacts to, primarily, Met496 in GluA2 imposed by the ethyl substituent of BPAM-97. These results add important information on binding affinities and thermodynamic details, and provide a new tool in the development of drugs against cognitive disorders.}}, author = {{Krintel, Christian and Frydenvang, Karla and Olsen, Lars and Kristensen, Maria T and de Barrios, Oriol and Naur, Peter and Francotte, Pierre and Pirotte, Bernard and Gajhede, Michael and Kastrup, Jette S}}, issn = {{0264-6021}}, language = {{eng}}, number = {{1}}, pages = {{173--178}}, publisher = {{Portland Press}}, series = {{Biochemical Journal}}, title = {{Thermodynamics and structural analysis of positive allosteric modulation of the ionotropic glutamate receptor GluA2}}, url = {{http://dx.doi.org/10.1042/BJ20111221}}, doi = {{10.1042/BJ20111221}}, volume = {{441}}, year = {{2012}}, }