The positive allosteric modulator BPAM344 and L-glutamate introduce an active-like structure of the ligand-binding domain of GluK2

Yasmin Bay; Mie Egeberg Jeppesen; Karla Frydenvang; Pierre Francotte; Bernard Pirotte; Darryl S. Pickering; Anders Skov Kristensen; Jette Sandholm Kastrup

doi:10.1002/1873-3468.14824

The positive allosteric modulator BPAM344 and L-glutamate introduce an active-like structure of the ligand-binding domain of GluK2

Yasmin Bay
, Mie Egeberg Jeppesen
, Karla Frydenvang
, Pierre Francotte
, Bernard Pirotte
, Darryl S. Pickering
, Anders Skov Kristensen
, Jette Sandholm Kastrup^*

^*Corresponding author for this work

Research output: Contribution to journal › Journal article › Research › peer-review

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Abstract

Kainate receptors belong to the family of ionotropic glutamate receptors and contribute to the majority of fast excitatory neurotransmission. Consequently, they also play a role in brain diseases. Therefore, understanding how these receptors can be modulated is of importance. Our study provides a crystal structure of the dimeric ligand-binding domain of the kainate receptor GluK2 in complex with L-glutamate and the small-molecule positive allosteric modulator, BPAM344, in an active-like conformation. The role of Thr535 and Gln786 in modulating GluK2 by BPAM344 was investigated using a calcium-sensitive fluorescence-based assay on transiently transfected cells expressing GluK2 and mutants hereof. This study may aid in the design of compounds targeting kainate receptors, expanding their potential as targets for the treatment of brain diseases.

Original language	English
Journal	FEBS Letters
Volume	598
Issue number	7
Pages (from-to)	743-757
ISSN	0014-5793
DOIs	https://doi.org/10.1002/1873-3468.14824
Publication status	Published - 2024

Bibliographical note

Publisher Copyright:
© 2024 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.

Funding

Beamline scientists at the MAX IV Laboratory, Lund, Sweden, are thanked for technical assistance. Linda Grønborg Dorvil is thanked for her technical support in the process of creating mutant constructs. Christophe Mulle is thanked for providing the GluK2() construct. Christina Kasper is thanked for providing the GluK2‐LBD (S1S2) construct and Heidi Peterson for technical assistance in expressing and purifying the GluK2‐LBD protein. The Independent Research Fund Denmark – Medical Sciences (YB, JSK) and Danscatt (YB, MEJ, KF, JSK) are acknowledged for financial support. We acknowledge the MAX IV Laboratory for time on Beamline BioMAX under proposal MX20200259. All funding sources had no part in the design of the study, the collection of data, or the analysis and interpretation, as well as no involvement in writing the manuscript or decision‐making of publication. Q

Keywords

BPAM344 binding mode
calcium-sensitive fluorescence-based assay
dimer interface
GluK2 mutations
positive allosteric modulation
X-ray crystallography

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title = "The positive allosteric modulator BPAM344 and L-glutamate introduce an active-like structure of the ligand-binding domain of GluK2",

abstract = "Kainate receptors belong to the family of ionotropic glutamate receptors and contribute to the majority of fast excitatory neurotransmission. Consequently, they also play a role in brain diseases. Therefore, understanding how these receptors can be modulated is of importance. Our study provides a crystal structure of the dimeric ligand-binding domain of the kainate receptor GluK2 in complex with L-glutamate and the small-molecule positive allosteric modulator, BPAM344, in an active-like conformation. The role of Thr535 and Gln786 in modulating GluK2 by BPAM344 was investigated using a calcium-sensitive fluorescence-based assay on transiently transfected cells expressing GluK2 and mutants hereof. This study may aid in the design of compounds targeting kainate receptors, expanding their potential as targets for the treatment of brain diseases.",

keywords = "BPAM344 binding mode, calcium-sensitive fluorescence-based assay, dimer interface, GluK2 mutations, positive allosteric modulation, X-ray crystallography",

author = "Yasmin Bay and \{Egeberg Jeppesen\}, Mie and Karla Frydenvang and Pierre Francotte and Bernard Pirotte and Pickering, \{Darryl S.\} and Kristensen, \{Anders Skov\} and Kastrup, \{Jette Sandholm\}",

note = "Publisher Copyright: {\textcopyright} 2024 The Authors. FEBS Letters published by John Wiley \& Sons Ltd on behalf of Federation of European Biochemical Societies.",

year = "2024",

doi = "10.1002/1873-3468.14824",

language = "English",

volume = "598",

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T1 - The positive allosteric modulator BPAM344 and L-glutamate introduce an active-like structure of the ligand-binding domain of GluK2

AU - Bay, Yasmin

AU - Egeberg Jeppesen, Mie

AU - Frydenvang, Karla

AU - Francotte, Pierre

AU - Pirotte, Bernard

AU - Pickering, Darryl S.

AU - Kristensen, Anders Skov

AU - Kastrup, Jette Sandholm

PY - 2024

Y1 - 2024

N2 - Kainate receptors belong to the family of ionotropic glutamate receptors and contribute to the majority of fast excitatory neurotransmission. Consequently, they also play a role in brain diseases. Therefore, understanding how these receptors can be modulated is of importance. Our study provides a crystal structure of the dimeric ligand-binding domain of the kainate receptor GluK2 in complex with L-glutamate and the small-molecule positive allosteric modulator, BPAM344, in an active-like conformation. The role of Thr535 and Gln786 in modulating GluK2 by BPAM344 was investigated using a calcium-sensitive fluorescence-based assay on transiently transfected cells expressing GluK2 and mutants hereof. This study may aid in the design of compounds targeting kainate receptors, expanding their potential as targets for the treatment of brain diseases.

AB - Kainate receptors belong to the family of ionotropic glutamate receptors and contribute to the majority of fast excitatory neurotransmission. Consequently, they also play a role in brain diseases. Therefore, understanding how these receptors can be modulated is of importance. Our study provides a crystal structure of the dimeric ligand-binding domain of the kainate receptor GluK2 in complex with L-glutamate and the small-molecule positive allosteric modulator, BPAM344, in an active-like conformation. The role of Thr535 and Gln786 in modulating GluK2 by BPAM344 was investigated using a calcium-sensitive fluorescence-based assay on transiently transfected cells expressing GluK2 and mutants hereof. This study may aid in the design of compounds targeting kainate receptors, expanding their potential as targets for the treatment of brain diseases.

KW - BPAM344 binding mode

KW - calcium-sensitive fluorescence-based assay

KW - dimer interface

KW - GluK2 mutations

KW - positive allosteric modulation

KW - X-ray crystallography

U2 - 10.1002/1873-3468.14824

DO - 10.1002/1873-3468.14824

M3 - Journal article

C2 - 38369668

AN - SCOPUS:85185969795

SN - 0014-5793

VL - 598

SP - 743

EP - 757

JO - FEBS Letters

JF - FEBS Letters

IS - 7

ER -