Gephyrin interacts with the glutamate receptor interacting protein 1 isoforms at GABAergic synapses

Wendou Yu, Erik I. Charych, David R. Serwanski, Rong Wen Li, Rashid Ali, Ben A. Bahr, Angel L. De Blas

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

We have previously shown that the glutamate receptor interacting protein 1 (GRIP1) splice forms GRIP1a/b and GRIP1c4-7 are present at the GABAergic post-synaptic complex. Nevertheless, the role that these GRIP1 protein isoforms play at the GABAergic post-synaptic complex is not known. We are now showing that GRIP1c4-7 and GRIP1a/b interact with gephyrin, the main post-synaptic scaffold protein of GABAergic and glycinergic synapses. Gephyrin coprecipitates with GRIP1c4-7 or GRIP1a/b from rat brain extracts and from extracts of human embryonic kidney 293 cells that have been cotransfected with gephyrin and one of the GRIP1 protein isoforms. Moreover, purified gephyrin binds to purified GRIP1c4-7 or GRIP1a/b, indicating that gephyrin directly interacts with the common region of these GRIP1 proteins, which includes PDZ domains 4-7. An engineered deletion construct of GRIP1a/b (GRIP1a4-7), which both contains the aforementioned common region and binds to gephyrin, targets to the post-synaptic GABAergic complex of transfected cultured hippocampal neurons. In these hippocampal cultures, endogenous gephyrin colocalizes with endogenous GRIP1c4-7 and GRIP1a/b in over 90% of the GABAergic synapses. Double-labeling electron microscopy immunogold reveals that in the rat brain GRIP1c4-7 and GRIP1a/b colocalize with gephyrin at the post-synaptic complex of individual synapses. These results indicate that GRIP1c4-7 and GRIP1a/b colocalize and interact with gephyrin at the GABAergic post-synaptic complex and suggest that this interaction plays a role in GABAergic synaptic function.

Original languageEnglish (US)
Pages (from-to)2300-2314
Number of pages15
JournalJournal of Neurochemistry
Volume105
Issue number6
DOIs
StatePublished - Jun 2008
Externally publishedYes

Fingerprint

Receptor-Interacting Protein Serine-Threonine Kinases
Glutamate Receptors
Synapses
Protein Isoforms
Rats
Brain
PDZ Domains
gephyrin
Scaffolds
Labeling
Electron microscopy
Neurons
Electron Microscopy
Proteins
Kidney

Keywords

  • Clustering
  • GABA synapse
  • GABA receptor
  • Gephyrin
  • Glutamate receptor interacting protein 1
  • PDZ domain

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Gephyrin interacts with the glutamate receptor interacting protein 1 isoforms at GABAergic synapses. / Yu, Wendou; Charych, Erik I.; Serwanski, David R.; Li, Rong Wen; Ali, Rashid; Bahr, Ben A.; De Blas, Angel L.

In: Journal of Neurochemistry, Vol. 105, No. 6, 06.2008, p. 2300-2314.

Research output: Contribution to journalArticle

Yu, Wendou ; Charych, Erik I. ; Serwanski, David R. ; Li, Rong Wen ; Ali, Rashid ; Bahr, Ben A. ; De Blas, Angel L. / Gephyrin interacts with the glutamate receptor interacting protein 1 isoforms at GABAergic synapses. In: Journal of Neurochemistry. 2008 ; Vol. 105, No. 6. pp. 2300-2314.
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AU - Ali, Rashid

AU - Bahr, Ben A.

AU - De Blas, Angel L.

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AB - We have previously shown that the glutamate receptor interacting protein 1 (GRIP1) splice forms GRIP1a/b and GRIP1c4-7 are present at the GABAergic post-synaptic complex. Nevertheless, the role that these GRIP1 protein isoforms play at the GABAergic post-synaptic complex is not known. We are now showing that GRIP1c4-7 and GRIP1a/b interact with gephyrin, the main post-synaptic scaffold protein of GABAergic and glycinergic synapses. Gephyrin coprecipitates with GRIP1c4-7 or GRIP1a/b from rat brain extracts and from extracts of human embryonic kidney 293 cells that have been cotransfected with gephyrin and one of the GRIP1 protein isoforms. Moreover, purified gephyrin binds to purified GRIP1c4-7 or GRIP1a/b, indicating that gephyrin directly interacts with the common region of these GRIP1 proteins, which includes PDZ domains 4-7. An engineered deletion construct of GRIP1a/b (GRIP1a4-7), which both contains the aforementioned common region and binds to gephyrin, targets to the post-synaptic GABAergic complex of transfected cultured hippocampal neurons. In these hippocampal cultures, endogenous gephyrin colocalizes with endogenous GRIP1c4-7 and GRIP1a/b in over 90% of the GABAergic synapses. Double-labeling electron microscopy immunogold reveals that in the rat brain GRIP1c4-7 and GRIP1a/b colocalize with gephyrin at the post-synaptic complex of individual synapses. These results indicate that GRIP1c4-7 and GRIP1a/b colocalize and interact with gephyrin at the GABAergic post-synaptic complex and suggest that this interaction plays a role in GABAergic synaptic function.

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