2anj: Difference between revisions

Revision as of 17:29, 21 February 2008

Crystal Structure of the Glur2 Ligand Binding Core (S1S2J-Y450W) Mutant in Complex With the Partial Agonist Kainic Acid at 2.1 A Resolution

OverviewOverview

Binding of an agonist to the 2-amino-3-(3-hydroxy-5-methyl-4-isoxazolyl)-propionic acid (AMPA) receptor family of the glutamate receptors (GluRs) results in rapid activation of an ion channel. Continuous application results in a non-desensitizing response for agonists like kainate, whereas most other agonists, such as the endogenous agonist (S)-glutamate, induce desensitization. We demonstrate that a highly conserved tyrosine, forming a wedge between the agonist and the N-terminal part of the bi-lobed ligand-binding site, plays a key role in the receptor kinetics as well as agonist potency and selectivity. The AMPA receptor GluR2, with mutations in Tyr-450, were expressed in Xenopus laevis oocytes and characterized in a two-electrode voltage clamp setup. The mutation GluR2(Y450A) renders the receptor highly kainate selective, and rapid application of kainate to outside-out patches induced strongly desensitizing currents. When Tyr-450 was substituted with the larger tryptophan, the (S)-glutamate desensitization is attenuated with a 10-fold increase in steady-state/peak currents (19% compared with 1.9% at the wild type). Furthermore, the tryptophan mutant was introduced into the GluR2-S1S2J ligand binding core construct and co-crystallized with kainate, and the 2.1-A x-ray structure revealed a slightly more closed ligand binding core as compared with the wild-type complex. Through genetic manipulations combined with structural and electrophysiological analysis, we report that mutations in position 450 invert the potency of two central agonists while concurrently strongly shaping the agonist efficacy and the desensitization kinetics of the AMPA receptor GluR2.

About this StructureAbout this Structure

2ANJ is a Single protein structure of sequence from Rattus norvegicus with as ligand. Full crystallographic information is available from OCA.

ReferenceReference

A binding site tyrosine shapes desensitization kinetics and agonist potency at GluR2. A mutagenic, kinetic, and crystallographic study., Holm MM, Naur P, Vestergaard B, Geballe MT, Gajhede M, Kastrup JS, Traynelis SF, Egebjerg J, J Biol Chem. 2005 Oct 21;280(42):35469-76. Epub 2005 Aug 15. PMID:16103115

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-[[Image:2anj.jpg|left|200px]]<br /><applet load="2anj" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:2anj.jpg|left|200px]]<br /><applet load="2anj" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="2anj, resolution 2.10&Aring;" />
 '''Crystal Structure of the Glur2 Ligand Binding Core (S1S2J-Y450W) Mutant in Complex With the Partial Agonist Kainic Acid at 2.1 A Resolution'''<br />
 ==Overview==
-Binding of an agonist to the, 2-amino-3-(3-hydroxy-5-methyl-4-isoxazolyl)-propionic acid (AMPA) receptor, family of the glutamate receptors (GluRs) results in rapid activation of, an ion channel. Continuous application results in a non-desensitizing, response for agonists like kainate, whereas most other agonists, such as, the endogenous agonist (S)-glutamate, induce desensitization. We, demonstrate that a highly conserved tyrosine, forming a wedge between the, agonist and the N-terminal part of the bi-lobed ligand-binding site, plays, a key role in the receptor kinetics as well as agonist potency and, selectivity. The AMPA receptor GluR2, with mutations in Tyr-450, were, expressed in Xenopus laevis oocytes and characterized in a two-electrode, voltage clamp setup. The mutation GluR2(Y450A) renders the receptor highly, kainate selective, and rapid application of kainate to outside-out patches, induced strongly desensitizing currents. When Tyr-450 was substituted with, the larger tryptophan, the (S)-glutamate desensitization is attenuated, with a 10-fold increase in steady-state/peak currents (19% compared with, 1.9% at the wild type). Furthermore, the tryptophan mutant was introduced, into the GluR2-S1S2J ligand binding core construct and co-crystallized, with kainate, and the 2.1-A x-ray structure revealed a slightly more, closed ligand binding core as compared with the wild-type complex. Through, genetic manipulations combined with structural and electrophysiological, analysis, we report that mutations in position 450 invert the potency of, two central agonists while concurrently strongly shaping the agonist, efficacy and the desensitization kinetics of the AMPA receptor GluR2.
+Binding of an agonist to the 2-amino-3-(3-hydroxy-5-methyl-4-isoxazolyl)-propionic acid (AMPA) receptor family of the glutamate receptors (GluRs) results in rapid activation of an ion channel. Continuous application results in a non-desensitizing response for agonists like kainate, whereas most other agonists, such as the endogenous agonist (S)-glutamate, induce desensitization. We demonstrate that a highly conserved tyrosine, forming a wedge between the agonist and the N-terminal part of the bi-lobed ligand-binding site, plays a key role in the receptor kinetics as well as agonist potency and selectivity. The AMPA receptor GluR2, with mutations in Tyr-450, were expressed in Xenopus laevis oocytes and characterized in a two-electrode voltage clamp setup. The mutation GluR2(Y450A) renders the receptor highly kainate selective, and rapid application of kainate to outside-out patches induced strongly desensitizing currents. When Tyr-450 was substituted with the larger tryptophan, the (S)-glutamate desensitization is attenuated with a 10-fold increase in steady-state/peak currents (19% compared with 1.9% at the wild type). Furthermore, the tryptophan mutant was introduced into the GluR2-S1S2J ligand binding core construct and co-crystallized with kainate, and the 2.1-A x-ray structure revealed a slightly more closed ligand binding core as compared with the wild-type complex. Through genetic manipulations combined with structural and electrophysiological analysis, we report that mutations in position 450 invert the potency of two central agonists while concurrently strongly shaping the agonist efficacy and the desensitization kinetics of the AMPA receptor GluR2.
 ==About this Structure==
-ANJ is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Rattus_norvegicus Rattus norvegicus] with KAI as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2ANJ OCA].
+ANJ is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Rattus_norvegicus Rattus norvegicus] with <scene name='pdbligand=KAI:'>KAI</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2ANJ OCA].
 ==Reference==
@@ Line 15: / Line 15: @@
 [[Category: Egebjerg, J.]]
 [[Category: Gajhede, M.]]
-[[Category: Geballe, M.T.]]
+[[Category: Geballe, M T.]]
-[[Category: Holm, M.M.]]
+[[Category: Holm, M M.]]
-[[Category: Kastrup, J.S.]]
+[[Category: Kastrup, J S.]]
 [[Category: Naur, P.]]
-[[Category: Traynelis, S.F.]]
+[[Category: Traynelis, S F.]]
 [[Category: Vestergaard, B.]]
 [[Category: KAI]]
@@ Line 27: / Line 27: @@
 [[Category: mutant]]
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